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Histology
1st year medical school



   Cambridge University
  by Christiane Riedinger
           2011
Aim of this presentation

               When starting to learn histology, I was looking for simple
               overviews presenting the key features of each tissue type
                              or tissue found in the body.
                Since I could not find that anywhere, I made it myself!

                  This presentation should be used with a standard
                 histology textbook (or the internet ;-)) showing you
                   slides/pictures of the structures described here.

                 Pictures of particular stains (part I) have been cited.




C.Riedinger
Contents

          1.   staining and fixing techniques

          2.   histological properties of human tissue types
                  2.1. connective tissue
                  2.2. muscle tissue
                  2.3. nervous tissue
                  2.4. epithelial tissue

          3.   histological properties of anatomical structures

          4.   ways to identify tissues/cells
                 • blood cells (connective tissue)
                 • ganglia (nervous tissue)
                 • components of the urinary system
C.Riedinger      • tissue types of the GI tract
1.     staining and fixing techniques




C.Riedinger
1.     staining and fixing techniques (2)




C.Riedinger
1.     staining and fixing techniques (3)




C.Riedinger
1.     staining and fixing techniques (4)




C.Riedinger
2.          tissue types: connective tissue
    Overview of tissue types:                                                                                                       C.Riedinger


        tissue                                                 layers/comp
                           function         cell types                                 appearance                    stains
         type                                                     onents
                        - binds           - fibroblasts        - collagen             types:                 - masson's trichrome
                        functional cell   (excrete ECM),       mostly I, II in        1. Loose (10-20% C)    (collagen)                 -
                        groupings         come from            cartilage, II in       2. Dense (40-50% C,    van gieson (collagen)
                        together          mesoderm             skin, vessels, IV in   tendons 90%)           - elastin stain (elastin)
                        - regulation      - adipocytes         epithelium of          3. elastic             - eosin (collagen, but not
                                          - chondroblasts      basement               lung, skin, bladder,   specific)
                                          - myofibroblasts     membranes              vessels,               - silver stain (reticulin)
                                          - immune cells:      - elastic fibres       change with age
                                          macrophages,         (stain poorly)         4. cartilage
                                          histiocytes, mast    - ground               70% ground
                                          cells, white blood   substance              substance
                                          cells                (= gel embedding       5. bone
                                                               collagen and           = cartilage with 70%
                                                               elastin)               salts
       connec-                                                                        6. fat
         tive                                                                         white or brown
                                                                                      7. blood
       (mesoderm)




C.Riedinger
2.          tissue types: muscle tissue
     Overview of tissue types:                                                                                                                   C.Riedinger


         tissue                                                  layers/comp
                             function        cell types                                   appearance                             stains
          type                                                      onents
                                                                                       - no striations, spindles
                                                                                                                         - masson's trichrome:
                                                                                       - 1 elongated centrally
                                                                                       located nucleus                   (muscle, connective
                                                                                       - irregularly branching           tissue)
                                                                                       fasciculi, can have ganglia
                                                                                       - shorter, often layered
                                           - unicellular:                              - fasciculi aren't in parallel
                                                                 - surrounded by
                                           myoepithelial cells                         - no myofibrils
                                                                 lamina
                                           (secretory glands),                         - caveolae
                                                                 - attached via link   - gap junctions
                                           pericytes (like
                                                                 proteins
                                           smooth muscle,
                                                                 - endomysium:         - intermediate
                                           surrounds blood                             - appears striated
                                                                 supp. tissue around
                                           vessels, called                             - 1-2 central nuclei
                                                                 each individual
                                           multiunit smooth                            - intracellular boundaries hard
                                                                 muscle fibre          to see
                           - contraction   muscle as each unit
       muscular                            functions
                                                                 - perimysium:         - cells appear continuous
                                                                                       (funct. Syncytium)
      (embryologically a                                         surrounds each
                                           individually),                              - branched ends
         subtype of                                              muscle cell bundle
                                           myofibroblasts                              - long, cylindrical
       connective from                                           = fascicle            - rich capillary network
                                           (contractile and
         mesoderm)                                               - epimysium:          - intercalated discs (Z)
                                           collagen, scarring)
                                                                 around groups of      - gap junctions
                                                                                       - no end plates, tendons!
                                                                 fasciculi, dense
                                           - multicellular:                            - diad (SR + T)
                                                                 collagenous sheeth
                                           smooth (lots of
                                                                 around whole
                                           cells function as                           - extremely elongated
                                                                 muscle
                                           single unit),                               - multinucleate (in
                                                                                       transverse section may not
                                           cardiac, skeletal
                                                                                       be seen)
                                                                                       - nucleus at periphery
                                                                                       - striated
                                                                                       - parallel fasciculi
                                                                                       - triad = SR terminal
                                                                                       cisterna + T-tubule




C.Riedinger
2.           tissue types: muscle tissue (2)
                                                                                                                                    C.Riedinger


     striated:

     light:         I-band containing Z-line (made of actin fibres)
     dark:          A-band containing myosin fibres

     H-band:        myosin-only region in A-band
     M-line:        middle of A-band

     T-tubules:
     T-tubules are at level of Z-bands (cardiac, amphibian skeletal)
     T-tubule at junction of A and I bands (skeletal)

     Red skeletal muscle: (aerobic, stains more strongly)
     rich in myoglobin, numeruous mitochondria, many capillaries
     White skeletal muscle: (anaerobic, stain is more pale)
     less myoglobin, fewer mitochondria, poorer blood supply

     cardiac:        junctions!
     intercalated discs:        black line perpendicular to length of fibre, parallelt o striations
     membrane-to-membrane contact in intercalated discs (only visible at EM resultion):
     1. Fascia adherens:        intermediate junction, anchors actin at terminal sarcomeres - mechanical connection
     2. desmosomes:             (macular adherens), attachment of intermediate filaments to cytoskeleton - mechanical
     3. Gap junctions:          (nexus), exchange/transmission of ions and small molecules from cell to cell - electrical
     result:                    functional syncytium!
     purkjinje fibres:          pacemaker cells, larger than cardiac muscle cells and sometimes binucleated
                                contain lots of mitochondria but less myofibrils (irregular), no T-tubules and intercalated discs
                                still have desmosomes and gap junctions, lots of glycogen (can stain for it specificlally!)

     smooth:
     dense bodies/plaques:        points of attachment for actin filaments
     caveolae:                    invaginations of the plasma membrane, help Ca2+ entry


     macula - spot (latin)


C.Riedinger
2.          tissue types: nervous tissue
Overview of tissue types:                                                                                                   C.Riedinger


    tissue                                                layers/comp
                       function          cell types                              appearance                     stains
     type                                                    onents
                     - electrically    - neurons          transverse:          - large cell body         - Nissl methylene blue
                     conduct signals   (multipolar,       - bundles of axons                             (rER)
                                       bipolar,           = fasciculi          - large, round,
                                       pseudounipolar)                         prominent but pale        - Sudan black (for LM)
                                       - glial cells      - endoneurium =      staining nucleus,         and osmium (for EM)
                                       (schwann cells,    around each nerve    dispersed chromatin       (myelin and lipids,
                                       oligodendrocyte,   fibre along with                               connective dissue)
                                       astrocytes,        myelin               - extensive basophilic
                                       satellite cells)                        cytoplasm
                                       - fibrocytes       - perineurium =
                                                          dense conn. tiss     - large and central
                                                          around bundles of    nucleolus
                                                          nerve fibres =       (transcriptional
                                                          fascicles            activity)

  nervous                                                 - epineurium =       - in longitudinal
 (from ectoderm?)                                         loose conn. tiss     section of nerve
                                                          around fascicles     trunks: zig-zaggy
                                                                               lines with round nuclei
                                                          Longitudinal:        (of schwann cells!!!!)
                                                          dendride, nodes of
                                                          ranvier              - abundant rER in
                                                          other: ganglia,      nucleus and dendrites
                                                          myelin, axon         (= Nissl substance
                                                          hillock, terminal    from Nissl staining
                                                                               RNA)

                                                                               - ganglia: cell bodies
                                                                               and/or synapses
2.           tissue types: epithelial tissue
      Overview of tissue types:                                                                                                C.Riedinger


          tissue                                                      layers/comp
                             function           cell types                                    appearance              stains
           type                                                          onents
                          - cover the body    - very closely          simple:
                          and line spaces     packed epithelial       = single layer
                          and tubes within    cells                   - squamous           - flat thin cells
                          it                  - subtype reflects                           difficult to distinguish
                                                                                           sometimes only nuclei
                          - protect           function
                                                                                           visible
                          - absorb            - goblet cells
                          - secrete           (mucus secreting)       - cuboidal           - round centrally
                                                                                           located nucleus, often
                          - skin, nephrons,   - hair cell (sensory)
                                                                                           polygonal
                          airways, glands,    - gustatory (taste
                          gut...              cell)                   - columnar           - tall, elongated cells
                                                                                           may be ciliated


                                              Even though not a       - pseudostratified   - mostly ciliated cells,
                                                                                           nuclei not in line
                                              cell type: epithelial
       epithelial                             tissue always
                                                                      stratified:
          (endoderm,                          contains a
                                              basement                = multiple layers
           ectoderm)
                                              membrane
                                              (lamina densa)          - squamous           - NEVER CILIATED!
                                              consisting of type      (wear & tear)
                                              IV collagen
                                                                      - cuboidal           - only top layer flat,
                                                                                           bottom layer cuboid

                                                                      - columnar

                                                                      - transitional       - intermediate betw.
                                                                                           Stratified cuboidal and
                                                                                           squamous. But all
                                                                                           layers have the same
                                                                                           shape!


C.Riedinger
3.            anatomical structures: vessels
 organs: vessels                                                                                                                                 C.Riedinger

                                                                        layers
   blood
                                                                                                                                                   other
  vessels                                    internal elastic                               external elastic
                   1. Tunica intima                                 2. Tunica media                             3. Tunica adventitia
                                                   lamina                                       lamina
              1a. Endothelium                   fenestrated     - smooth muscle                                - supporting tissue:
              (1 flat layer, cells difficult layer of elastin   - collagen                                     collagen
              to distinguish in LM, often separating 1.         - elastin                     less defined     - contains innervation
              see only nuclei)               and 2.. In very    - quite thick compared to                      and blood supply (for
                                                                                                layer of
              1b. Basement                     large elastic    intima                                         very large vessels, vasa
  general                                                                                       elasting
              membrane                       vessels hard to                                                   vasorum)
                                                                                             separating 2.
              1c. Connective tissue           see as media                                                     - in continuation with
                                                                                                 and 3.
                                              has so many                                                      surrounding tissue
                                                  layers of
                                                   elastin.
                                                                            +
                                                                  very broad, contains
                                                                 concentrically arranged
 arteries,
                           +                        +             layers of elastin with           +                      +
  elastic
                                                                  some smooth muscle
                                                                 between layers, elastin
                                                                   decreases with age
                                                                            +
 arteries,                                                          circumferentially
                           +                        +                                              +                      +
 muscular                                                           arranged smooth
                                                                         muscle
                                                                           +                                              +
                           +
 arteriole                                          +            (almost entirely smooth           -                 (merges with           <0.3mm diam
                         (thin)
                                                                         muscle)                                  surrounding tissue)
                                                                                                                                              overall much
                           +                                                                                              +                   thinner wall
    vein                                             -                      +                      -
                         (thin)                                                                                    (most prominent)           compared to
                                                                          (thin)
                                                                                                                                                 lumen
   lymph                          like veins but no erys in lumen, few leucocytes and precipitaed lymp protein (artifact of preparation!)
                                                                                                                                             continuous, fenestrated
                          +                                                                                                                 (windows bridged by thin
capillaries        (only 1a and 1b)                  -                      -                      -                       -              diaphragm) or sinusuidal with
                                                                                                                                                   proper gaps

  venule                    +                       -                    -                      -                    +
nuclei of endothelial cells are elongated in direction of vessel, smooth muscle nuclei are elongated circumferentially
3.          anatomical structures: respiratory system
organs: respiratory system (lung)                                                                                                              C.Riedinger



                                               1. mucosa?
  compart-
                                                                                          2. submucosa            3. cartilage            stain
    ment
               1a. epithelium              1b. Lamina propria 1c. Smooth Muscle

                                                                                        - numerous         - C-shaped hyaline          - H&E
               - pseudostratified
                                                                                        seromucinous       cartilage
               columnar ciliated           - loose connective
                                                                                        glands             - with layers of
               - many mucus                tissue                                                          fibroelastic tissue
                                                                                        - serous cells stain
trachea        secreting goblet                                            -
                                           - many blood                                                    between cart. rings
               cells                                                                    strongly
                                           vessels                                                         - submucosa
               - unusually thick                                                        - mucous cells stain
                                                                                        poorly             merges with its
               basement
                                                                                                           perichondrium
               - pseudostratified                                                       - fewer            - flatter,
               columnar ciliated                                                        seromuceous glands interconnected
               (less tall, smaller)        - more dense                                                    plates of cartilage
bronchus       - fewer goblet cells        - more elastic                  +                               rather than rings
                                                                                                           - not C-shaped



               - simple columnar
                                                                                        -seems to be there              -
               ciliated
               - <1mm diam.                -very thin - is it                           but much less
               - smaller bronchioles
                                                                           +            glands and thinner        thin-walled
                                           there?
bronchioles                                                                                                    pulmonary artery
               cuboidal
                                                                  prominent feature!
               - less to no goblet                                                                              branches can lie
               cells, but Clara cells!                                                                         next to bronchiole
               (= resp. bronchiole)
               - lined with pneumo-        - alveolar ducts: smooth muscle cells, collagen and elastic fibres
               cyte type I cells (40%      - alveolar septum (wall): alveolar capillaries and sparse network of elastin and collagen
               covering 90%)
                                           with pneumocytes of the walls of the two adjacent alveoli next to it
               - can only see nuclei
               - 60/10% pneumocyte         - septum also contains few fibroblasts
alveoli
               type II (cuboidal, much     - elastin and collagen condense around alveolar openings to form supporting network
               CP, secrete surfactant)     for lung parenchyma (parenchyme = bulk of a substance = lung material)
               - endothelial cell on the
                                           - 8um openings in septum: alveolar pores (of Kohn) for air exachange
               blood side
                                           - also alveolar macrophages (dust cells) with thin flattened, even nucleus
3.             anatomical structures: urinary system
   organs: urinary system (kidneys)                                                                                                         C.Riedinger
compartment                    epithelium                           other cells              features             (Wheater's p. 318, 320,325)

                               simple squamous endothelium          1. juxtaglomerular
     afferent arteriole                                                                      - modified smooth muscle cells
                                                                    cells
                               simple squamous endothelium          - podocytes              - 1* and 2* foot processes
                               (fenestrated)                        - mesangial cells        - embrace capillary loops
                                                                                             - filtration barrier: 1. Endothelial cells, 2. Basement
                                                                     2. extraglomerular
                                                                                             membrane, 3. podocytes
                                                                    mesangial cells
                                                                                             - contractive cells, phagocytotic (can reduce GFR)
                  glomerulus
                                          JUXTAGLOMERULAR APPARATUS                          - surround glomerular capillaries
renal                                         formed by comparments of                       - mesangium = supportive tissue similar to basement
corpuscle                                  glomerulus, afferent arteriole and                membrane, cytoplasm very stained
                                                                                             - flat, elongated continuous with glomerular mesang. cells
                                                distal convoluted tubule
                                                                                             - conical mass, cytoplasmic processes
                               simple squamous epithelium                                    - invaginated sphere
             bowman's
                                                                                -            - visceral and parietal layer
             capsule
                                                                                             (but where is visceral layer? Can't see it on EM)
                               simple cuboidal epithelium                                    - brush border (aids reabsorbtion)
proximal convoluted tubule
                                                                                  -          - many mitochondria, endocytotic vesicles and
(PCT)
                                                                                             lysosomes - FUZZY LUMEN!!!
thin descending and thin       simple squamous epithelium
                                                                                -
ascending loop of henle
thick ascending loop of        simple cuboidal epithelium (low)
                                                                                -
henle
                                                                    3. macula densa          -   NO brush border!
                               simple cuboidal epithelium                                    -   smaller cells, stain less intensely
                                                                                                                                     - CLEAR LUMEN!!!
                                                                    (where in contact with
                                                                                             -   nucleus protrudes into lumen
distal convoluted tubule                                            glomerulus)
                                                                                             -   cell volume smaller
(DCT)
                                                                                             -   specialised epithelial cells
                                                                                             -   closely packed, taller, thin basement membrane
                                                                                             -   located on side of DCT that faces corpuscle
                               simple cuboidal epithelium                                    - wider than CDT
collecting tubules (CT)                                                         -
                                                                                             - less regular in shape
                               simple columnar epithelium           - principal cells        -   large diameter - pale cytoplasm, few organelles
collecting duct (CD)                                                - a-intercalated cells   -   pale stained     - short microvilli
                                                                                             -   darker cytoplasm, many mitos, vesicles (H+)
                               stratified transitional epithelium                            -   3-6 layers
ureter, bladder                                                                 -            -   thick luminal surface
                                                                                             -   impermeable to urine/water
3.          anatomical structures: glands
   organs: glands                                                                                                                                        C.Riedinger

type of gland       arrangement            main component                   ducts                          features                              other
                                         acini (end pieces) =
                                                                     intercalated ducts
                                          clusters of mucus                              mucus secreting: bigger, sain pale             lots of vessels, nerves,
                 lobules separated by                                leading to striated
                                        secreting cells, serous                             with nuclei on side. Serous              (parasympathetic) ganglions,
Salivary         septa, surrounded by                                 ducts, stain red.
                                      secreting cells, or a mix of                       secreting: smaller, stain strongly,          extretory ducts, connective
                        capsule                                     Simple cuboidal epi,
                                       both. Duct system more                                pyramidal/cuboidal cells                  tissue, lymphatic vessels
                                                                   central round nucleus
                                              prominent

                                        acini (exclusively serous       intercalated ducts
                                                                                                   arranged circularly with         abundant blood supply, network
                                           with central nuclei,        (difficult to ID with
                                                                                                 lateral nuclei (apex towards         of arterioles.      in ducts:
                                           surrounded by fine             LM), leading to
                                                                                                   inside, nucleus basal), tiny     cells change from squamous or
                                         network of supporting           intralobular and
                  lobules separated by                                                          lumen, sometimes centroacinar       cuboidal epithelium to stratified
                                            tissue containing        interlobular ducts (big
                loose supporting tissue                                                                        cells                     cuboidal in large ducts
                                                sinusoids)                    lumen)
                      surrounded by
Pancreas          collagenous capsule.                                                                                              immunological stain for glucagen
                Exocrine (80-85%) and                                                           smaller, scattered pale staining     (alpha-cells, smaller) or insulin
                   endocrine (1-2%)                                                                blobs of varying size, cells      (beta-cells, stain with aldehyde
                         features!        islets of langerhans                  -                contained smaller than acinar      fuchsin) reveals that glucagon is
                                                                                               cells, evenly distributed cells with     produced in the periphery
                                                                                                    evenly distributed nuclei          whereas insulin is produced
                                                                                                                                                 centrally


                                                                                                 polyhedral cells with round
                                                                                                  nuclei, some binucleated,
                                         hepatocytes and sinusoid                              arranged into branching sheaths
                                                arterioles                                     of 1 cell thickness, separated by
                                                                                               sinusoids which appear as empty       macrophages (Kupffer cells)
                                                                                                              spaces                present in sinusoids to remove
                polygonal lobules with
                                                                   canaliculi with microvilli                                         debris, sinusoids have gaps
                  thin boundaries of
Liver                                                              that run countercurrent                                           between endothelial cells to
                collagenous supporting
                                       portal tracts/triads: entry     to the sinusoids                                              promote exchange of plasma
                        tissue
                                           site of blood from                                 thin-walled veins, thicker walled          components with the
                                         terminal branches of                                   arteries, darkly staining bile          hepatocytes (see EM).
                                        portal vein and hepatic                                ducts. Also contains lymphatic
                                        artery, leads to central                                 tissue/ducts which is often
                                         vein, exit of bile duct                                          collapsed
                                              (canaliculus).
3.             anatomical structures: glands (2)
   organs: glands                                                                                                                                               C.Riedinger

type of gland       arrangement               components                   products                          features                                   other
                                         follicles, contain thyroid                               morphology ~ activity: resting                  hormones bound to
                                            hormones stored in         tri-iodothyronine, 4-     thyroid follicular cells flattened,         thyroglobulin, a glycoprotein,
                                           homogenous colloids,             iodothyronine          lots of colloid, active thyroid          when stored. Thyroid gland is
                                         lined with single layer of          (=thyroxine)         follicular cells large, columnar,        unique in storing lots of hormone
   Thyroid            lobulated          cuboidal follicular cells                                   basal nucleus, less colloid                    when inactive!

                                                                                                scattered, lumps or single cells,
                                                                                                                                            endoneurocrine, derived from
                                           parafollicular cells             calcitonin           near fenestrated capillaries for
                                                                                                                                                 neural crest cells?
                                                                                                hormones to enter blood stream


                poorly defined lobules                                                           large, round nuclei, resting cells have
                                          chief/principal cells                                 pale cytoplasm, prominent golgi, rER,
                 + septa contained in                                          PTH
                                             (most common)                                          secretory granules. When active
                the capsule of thyroid                                                                                              glandular elements can be
                                                                                                smaller, more rER, stain more strongly
                gland (septa [blue] =                                                                                            intermixed with adipose cells, in
 Parathyroid
                extensions of capsule                                                                                               age becomes infiltrated by
                      containing                                                                eosinophilic cytoplasm, numerous          lymphocytes
                    neurovascular         oxyphil cells (minor
                                                                            unknown                 mitochondria, larger than
                     structures)             component)
                                                                                                          principal cells


                                         glandular epithelium,
                                            intimate vascular                                   cords or clumps of cells, sinusoid
                                                                                                                                               *50% somatrophs (GH)
                                             connections with                                   capillaries, collagen and reticulin
Anteroir                                                                                                                                      20% corcitutrophs (ACTH)
                                              hypothalamus.            GH, ACTH, LH, FSH,           network, chromophobes:
pituitary               blob                                                                                                                 20% lactrotrophs (prolactin)
                                            chromophobe and           prolactin, thyrotrophin        smallest, few granules,
(adeno)                                                                                                                                      5% gonadotrophs (LH, FSH)
                                           chromophil cells =                                       chromophils: acidophil or
                                                                                                                                            5% thyrotrophs (thyrotrophin)
                                         ~troph cells with lots of                                           basophil.
                                                granules*

                                                                                                      axons: lots of granules,
                                                                                                      accumulate in distended
                                                                                                  terminations = Herring bodies,
Posterior                                non-myelinated axons,
                     connects to                                                                     granules contain hormone                   cell bodies of axons in
pituitary                                pituicytes (speciallised         ADH, oxytocin
                hypothalamus via stalk                                                          precursors generated in cell body,                   hypothalamus
(neuro)                                         glial cells)
                                                                                                 final hormone generated during
                                                                                                    transport. Pituicyte EM: few
                                                                                                              granules
3.   anatomical structures: GI tract
                                       C.Riedinger
4.         identify: blood cells (connective tissue)

        BLOOD                                                        Wheater's:     table page 64
        1. red blood cells:
        - total absence of organelles
        - no nucleus
        - flattened disc with elevated circumference
        - reticulocytes (= precursors, <1% of circulating erys) have some residual nuclear material
        - very young cells some rER and mitochondria

        2. white blood cells: (1/1000 blood cells)
        5 types, named based on staining properties of granules
        granulocytes:               Neutrophils       do not stain in humans                                  60%
                                    eosinophils       pick up eosin and therefore stain orange                 3%
                                    basophils         pick up azures and therefore stain blue, rarest cells    1%
                                    single multilobed nuclei (polymorphonuclear)
                                    originally believed to be polynuclear
        mononuclear leucocytes:     lymphocytes clear cytoplasm, rounded nucleus                              34%
        (agranular)                 monocytes         large, indented curved nucleus                           4%
                                    non-lobulated nuclei
                                    agranolucytes

        3. platelets:
        - small
        - non-nucleated
        - round or oval, biconvex
        - cytoplasm purple stained
        - granules = 20% of platelet volume
        - many organelles




C.Riedinger
4.      identify: blood cells (connective tissue) (2)




C.Riedinger
4.          identify: type of ganglia (nervous tissue)

                                                                                                                                        C.Riedinger


how to distinguish ganglia
cell bodies are large with smaller supporting cells around it
symp + parasymp:           contain synapses                                           (stellar ganglion = largest symp ganglion)
sensory:                   just contain cell bodies

appearance
sensory ganglion:         many and larger nuclei of satellite (supporting cells), form neat circle around cell body, even larger cell bodies
                          pseudounipolar neurons!
sympathetic ganglion:     smaller and more scattered satellites, smaller cell bodies
                          more space between cell bodies as axons and dendrites have to pass through!
                          same basic structure as sensory ganglia
parasymp. Ganglion:       near target organ! Islands of connective tissue with blood vessels, nerves and ducts
                          nerve cell bodies lie within nerve trunk, are surrounded by support cells,
                          less satellites, smaller cell bodies
                          large cell bodies and axons

nerves in longitudinal section
zig-zaggy strands with nuclei of schwann cells visible
each myelin producing schwann cell covers ca. 1mm of the nerve fibre
in-between: nodes of ranvier
often stained black with pink connective tissue in-between individual myelinated fibres
4.        identify: GI tract
                                      - looks more structured than
                                      stomach                                                                               C.Riedin
organs: alimentary system (gut)
                                      - mucin stains more blue than
                                      cyan
Wheater's:                      page 286, 287, for glands 97


How to distinguish different parts of the gut:

Are there villi?
          * Yes                Small intestine: Duodenum/Jejunum/Ileum
                               Are there brunner's glands?
                                                    * Yes              Duodenum
                                                    * No               Jejunum/Ileum
                                                                       Are there peyer's patches?
                                                                                         * Yes                    Ileum
           * No                Oesophagus/Stomach/Colon                                  * No                     Jejunum
                               Are there glands?
                                                    * Yes              Stomach/Colon
                                                                       What do the glands look like?
                                                                                         Straight and beautiful   Colon
                                                                                         Thick very thick layer
                                                                                         underneath ducts, less   Stomach
                                                                                         ordered
                                                    * No               Stratified squamout epithelium?            Oesophagus
4.           identify: urinary system


   organs: urinary system (kidneys)                                                                                             C.Riedinger

adrenal gland

egulfed in dense supporting tissue that extends into gland to support secretory cells

               zona glomerulosa 5-10%
                                 secretes mineralocorticoids (e.g. aldosterone)
                                 whorls of cells and capillaries
               zona fasciculata 75%
                                 narrow cords of large cels
                                 sinusoid capillaries
cortex:                          rich in sER and lipids
                                 "foamy"
                                 secretes glucocorticoids (e.g. cortisol)
               zona reticulosa   irregular network of branching cords
                                 numerous capillaries of wide diameter
                                 smaller cells than other two layers
                                 secretes androgenic steroids
medulla:       chromaffin cells clumps and cords of cells
                                 surrounded by fine supporting tissue
                                 large nucleus (stains blue)
                                 basophilic cytoplasm
               secretes catecholamines (e.g. (nor)adrenaline)

steroid secreting cells:         many mitochondria with unusual tubular cristae
                                 sER
                                 lipid droplets (if secreting cholesterol, in cortex)
                                 membrane-bound granules (if secreting catecholamines in medulla, but those are not steroids)
The End.

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Histology 1st year medschool

  • 1. Histology 1st year medical school Cambridge University by Christiane Riedinger 2011
  • 2. Aim of this presentation When starting to learn histology, I was looking for simple overviews presenting the key features of each tissue type or tissue found in the body. Since I could not find that anywhere, I made it myself! This presentation should be used with a standard histology textbook (or the internet ;-)) showing you slides/pictures of the structures described here. Pictures of particular stains (part I) have been cited. C.Riedinger
  • 3. Contents 1. staining and fixing techniques 2. histological properties of human tissue types 2.1. connective tissue 2.2. muscle tissue 2.3. nervous tissue 2.4. epithelial tissue 3. histological properties of anatomical structures 4. ways to identify tissues/cells • blood cells (connective tissue) • ganglia (nervous tissue) • components of the urinary system C.Riedinger • tissue types of the GI tract
  • 4. 1. staining and fixing techniques C.Riedinger
  • 5. 1. staining and fixing techniques (2) C.Riedinger
  • 6. 1. staining and fixing techniques (3) C.Riedinger
  • 7. 1. staining and fixing techniques (4) C.Riedinger
  • 8. 2. tissue types: connective tissue Overview of tissue types: C.Riedinger tissue layers/comp function cell types appearance stains type onents - binds - fibroblasts - collagen types: - masson's trichrome functional cell (excrete ECM), mostly I, II in 1. Loose (10-20% C) (collagen) - groupings come from cartilage, II in 2. Dense (40-50% C, van gieson (collagen) together mesoderm skin, vessels, IV in tendons 90%) - elastin stain (elastin) - regulation - adipocytes epithelium of 3. elastic - eosin (collagen, but not - chondroblasts basement lung, skin, bladder, specific) - myofibroblasts membranes vessels, - silver stain (reticulin) - immune cells: - elastic fibres change with age macrophages, (stain poorly) 4. cartilage histiocytes, mast - ground 70% ground cells, white blood substance substance cells (= gel embedding 5. bone collagen and = cartilage with 70% elastin) salts connec- 6. fat tive white or brown 7. blood (mesoderm) C.Riedinger
  • 9. 2. tissue types: muscle tissue Overview of tissue types: C.Riedinger tissue layers/comp function cell types appearance stains type onents - no striations, spindles - masson's trichrome: - 1 elongated centrally located nucleus (muscle, connective - irregularly branching tissue) fasciculi, can have ganglia - shorter, often layered - unicellular: - fasciculi aren't in parallel - surrounded by myoepithelial cells - no myofibrils lamina (secretory glands), - caveolae - attached via link - gap junctions pericytes (like proteins smooth muscle, - endomysium: - intermediate surrounds blood - appears striated supp. tissue around vessels, called - 1-2 central nuclei each individual multiunit smooth - intracellular boundaries hard muscle fibre to see - contraction muscle as each unit muscular functions - perimysium: - cells appear continuous (funct. Syncytium) (embryologically a surrounds each individually), - branched ends subtype of muscle cell bundle myofibroblasts - long, cylindrical connective from = fascicle - rich capillary network (contractile and mesoderm) - epimysium: - intercalated discs (Z) collagen, scarring) around groups of - gap junctions - no end plates, tendons! fasciculi, dense - multicellular: - diad (SR + T) collagenous sheeth smooth (lots of around whole cells function as - extremely elongated muscle single unit), - multinucleate (in transverse section may not cardiac, skeletal be seen) - nucleus at periphery - striated - parallel fasciculi - triad = SR terminal cisterna + T-tubule C.Riedinger
  • 10. 2. tissue types: muscle tissue (2) C.Riedinger striated: light: I-band containing Z-line (made of actin fibres) dark: A-band containing myosin fibres H-band: myosin-only region in A-band M-line: middle of A-band T-tubules: T-tubules are at level of Z-bands (cardiac, amphibian skeletal) T-tubule at junction of A and I bands (skeletal) Red skeletal muscle: (aerobic, stains more strongly) rich in myoglobin, numeruous mitochondria, many capillaries White skeletal muscle: (anaerobic, stain is more pale) less myoglobin, fewer mitochondria, poorer blood supply cardiac: junctions! intercalated discs: black line perpendicular to length of fibre, parallelt o striations membrane-to-membrane contact in intercalated discs (only visible at EM resultion): 1. Fascia adherens: intermediate junction, anchors actin at terminal sarcomeres - mechanical connection 2. desmosomes: (macular adherens), attachment of intermediate filaments to cytoskeleton - mechanical 3. Gap junctions: (nexus), exchange/transmission of ions and small molecules from cell to cell - electrical result: functional syncytium! purkjinje fibres: pacemaker cells, larger than cardiac muscle cells and sometimes binucleated contain lots of mitochondria but less myofibrils (irregular), no T-tubules and intercalated discs still have desmosomes and gap junctions, lots of glycogen (can stain for it specificlally!) smooth: dense bodies/plaques: points of attachment for actin filaments caveolae: invaginations of the plasma membrane, help Ca2+ entry macula - spot (latin) C.Riedinger
  • 11. 2. tissue types: nervous tissue Overview of tissue types: C.Riedinger tissue layers/comp function cell types appearance stains type onents - electrically - neurons transverse: - large cell body - Nissl methylene blue conduct signals (multipolar, - bundles of axons (rER) bipolar, = fasciculi - large, round, pseudounipolar) prominent but pale - Sudan black (for LM) - glial cells - endoneurium = staining nucleus, and osmium (for EM) (schwann cells, around each nerve dispersed chromatin (myelin and lipids, oligodendrocyte, fibre along with connective dissue) astrocytes, myelin - extensive basophilic satellite cells) cytoplasm - fibrocytes - perineurium = dense conn. tiss - large and central around bundles of nucleolus nerve fibres = (transcriptional fascicles activity) nervous - epineurium = - in longitudinal (from ectoderm?) loose conn. tiss section of nerve around fascicles trunks: zig-zaggy lines with round nuclei Longitudinal: (of schwann cells!!!!) dendride, nodes of ranvier - abundant rER in other: ganglia, nucleus and dendrites myelin, axon (= Nissl substance hillock, terminal from Nissl staining RNA) - ganglia: cell bodies and/or synapses
  • 12. 2. tissue types: epithelial tissue Overview of tissue types: C.Riedinger tissue layers/comp function cell types appearance stains type onents - cover the body - very closely simple: and line spaces packed epithelial = single layer and tubes within cells - squamous - flat thin cells it - subtype reflects difficult to distinguish sometimes only nuclei - protect function visible - absorb - goblet cells - secrete (mucus secreting) - cuboidal - round centrally located nucleus, often - skin, nephrons, - hair cell (sensory) polygonal airways, glands, - gustatory (taste gut... cell) - columnar - tall, elongated cells may be ciliated Even though not a - pseudostratified - mostly ciliated cells, nuclei not in line cell type: epithelial epithelial tissue always stratified: (endoderm, contains a basement = multiple layers ectoderm) membrane (lamina densa) - squamous - NEVER CILIATED! consisting of type (wear & tear) IV collagen - cuboidal - only top layer flat, bottom layer cuboid - columnar - transitional - intermediate betw. Stratified cuboidal and squamous. But all layers have the same shape! C.Riedinger
  • 13. 3. anatomical structures: vessels organs: vessels C.Riedinger layers blood other vessels internal elastic external elastic 1. Tunica intima 2. Tunica media 3. Tunica adventitia lamina lamina 1a. Endothelium fenestrated - smooth muscle - supporting tissue: (1 flat layer, cells difficult layer of elastin - collagen collagen to distinguish in LM, often separating 1. - elastin less defined - contains innervation see only nuclei) and 2.. In very - quite thick compared to and blood supply (for layer of 1b. Basement large elastic intima very large vessels, vasa general elasting membrane vessels hard to vasorum) separating 2. 1c. Connective tissue see as media - in continuation with and 3. has so many surrounding tissue layers of elastin. + very broad, contains concentrically arranged arteries, + + layers of elastin with + + elastic some smooth muscle between layers, elastin decreases with age + arteries, circumferentially + + + + muscular arranged smooth muscle + + + arteriole + (almost entirely smooth - (merges with <0.3mm diam (thin) muscle) surrounding tissue) overall much + + thinner wall vein - + - (thin) (most prominent) compared to (thin) lumen lymph like veins but no erys in lumen, few leucocytes and precipitaed lymp protein (artifact of preparation!) continuous, fenestrated + (windows bridged by thin capillaries (only 1a and 1b) - - - - diaphragm) or sinusuidal with proper gaps venule + - - - + nuclei of endothelial cells are elongated in direction of vessel, smooth muscle nuclei are elongated circumferentially
  • 14. 3. anatomical structures: respiratory system organs: respiratory system (lung) C.Riedinger 1. mucosa? compart- 2. submucosa 3. cartilage stain ment 1a. epithelium 1b. Lamina propria 1c. Smooth Muscle - numerous - C-shaped hyaline - H&E - pseudostratified seromucinous cartilage columnar ciliated - loose connective glands - with layers of - many mucus tissue fibroelastic tissue - serous cells stain trachea secreting goblet - - many blood between cart. rings cells strongly vessels - submucosa - unusually thick - mucous cells stain poorly merges with its basement perichondrium - pseudostratified - fewer - flatter, columnar ciliated seromuceous glands interconnected (less tall, smaller) - more dense plates of cartilage bronchus - fewer goblet cells - more elastic + rather than rings - not C-shaped - simple columnar -seems to be there - ciliated - <1mm diam. -very thin - is it but much less - smaller bronchioles + glands and thinner thin-walled there? bronchioles pulmonary artery cuboidal prominent feature! - less to no goblet branches can lie cells, but Clara cells! next to bronchiole (= resp. bronchiole) - lined with pneumo- - alveolar ducts: smooth muscle cells, collagen and elastic fibres cyte type I cells (40% - alveolar septum (wall): alveolar capillaries and sparse network of elastin and collagen covering 90%) with pneumocytes of the walls of the two adjacent alveoli next to it - can only see nuclei - 60/10% pneumocyte - septum also contains few fibroblasts alveoli type II (cuboidal, much - elastin and collagen condense around alveolar openings to form supporting network CP, secrete surfactant) for lung parenchyma (parenchyme = bulk of a substance = lung material) - endothelial cell on the - 8um openings in septum: alveolar pores (of Kohn) for air exachange blood side - also alveolar macrophages (dust cells) with thin flattened, even nucleus
  • 15. 3. anatomical structures: urinary system organs: urinary system (kidneys) C.Riedinger compartment epithelium other cells features (Wheater's p. 318, 320,325) simple squamous endothelium 1. juxtaglomerular afferent arteriole - modified smooth muscle cells cells simple squamous endothelium - podocytes - 1* and 2* foot processes (fenestrated) - mesangial cells - embrace capillary loops - filtration barrier: 1. Endothelial cells, 2. Basement 2. extraglomerular membrane, 3. podocytes mesangial cells - contractive cells, phagocytotic (can reduce GFR) glomerulus JUXTAGLOMERULAR APPARATUS - surround glomerular capillaries renal formed by comparments of - mesangium = supportive tissue similar to basement corpuscle glomerulus, afferent arteriole and membrane, cytoplasm very stained - flat, elongated continuous with glomerular mesang. cells distal convoluted tubule - conical mass, cytoplasmic processes simple squamous epithelium - invaginated sphere bowman's - - visceral and parietal layer capsule (but where is visceral layer? Can't see it on EM) simple cuboidal epithelium - brush border (aids reabsorbtion) proximal convoluted tubule - - many mitochondria, endocytotic vesicles and (PCT) lysosomes - FUZZY LUMEN!!! thin descending and thin simple squamous epithelium - ascending loop of henle thick ascending loop of simple cuboidal epithelium (low) - henle 3. macula densa - NO brush border! simple cuboidal epithelium - smaller cells, stain less intensely - CLEAR LUMEN!!! (where in contact with - nucleus protrudes into lumen distal convoluted tubule glomerulus) - cell volume smaller (DCT) - specialised epithelial cells - closely packed, taller, thin basement membrane - located on side of DCT that faces corpuscle simple cuboidal epithelium - wider than CDT collecting tubules (CT) - - less regular in shape simple columnar epithelium - principal cells - large diameter - pale cytoplasm, few organelles collecting duct (CD) - a-intercalated cells - pale stained - short microvilli - darker cytoplasm, many mitos, vesicles (H+) stratified transitional epithelium - 3-6 layers ureter, bladder - - thick luminal surface - impermeable to urine/water
  • 16. 3. anatomical structures: glands organs: glands C.Riedinger type of gland arrangement main component ducts features other acini (end pieces) = intercalated ducts clusters of mucus mucus secreting: bigger, sain pale lots of vessels, nerves, lobules separated by leading to striated secreting cells, serous with nuclei on side. Serous (parasympathetic) ganglions, Salivary septa, surrounded by ducts, stain red. secreting cells, or a mix of secreting: smaller, stain strongly, extretory ducts, connective capsule Simple cuboidal epi, both. Duct system more pyramidal/cuboidal cells tissue, lymphatic vessels central round nucleus prominent acini (exclusively serous intercalated ducts arranged circularly with abundant blood supply, network with central nuclei, (difficult to ID with lateral nuclei (apex towards of arterioles. in ducts: surrounded by fine LM), leading to inside, nucleus basal), tiny cells change from squamous or network of supporting intralobular and lobules separated by lumen, sometimes centroacinar cuboidal epithelium to stratified tissue containing interlobular ducts (big loose supporting tissue cells cuboidal in large ducts sinusoids) lumen) surrounded by Pancreas collagenous capsule. immunological stain for glucagen Exocrine (80-85%) and smaller, scattered pale staining (alpha-cells, smaller) or insulin endocrine (1-2%) blobs of varying size, cells (beta-cells, stain with aldehyde features! islets of langerhans - contained smaller than acinar fuchsin) reveals that glucagon is cells, evenly distributed cells with produced in the periphery evenly distributed nuclei whereas insulin is produced centrally polyhedral cells with round nuclei, some binucleated, hepatocytes and sinusoid arranged into branching sheaths arterioles of 1 cell thickness, separated by sinusoids which appear as empty macrophages (Kupffer cells) spaces present in sinusoids to remove polygonal lobules with canaliculi with microvilli debris, sinusoids have gaps thin boundaries of Liver that run countercurrent between endothelial cells to collagenous supporting portal tracts/triads: entry to the sinusoids promote exchange of plasma tissue site of blood from thin-walled veins, thicker walled components with the terminal branches of arteries, darkly staining bile hepatocytes (see EM). portal vein and hepatic ducts. Also contains lymphatic artery, leads to central tissue/ducts which is often vein, exit of bile duct collapsed (canaliculus).
  • 17. 3. anatomical structures: glands (2) organs: glands C.Riedinger type of gland arrangement components products features other follicles, contain thyroid morphology ~ activity: resting hormones bound to hormones stored in tri-iodothyronine, 4- thyroid follicular cells flattened, thyroglobulin, a glycoprotein, homogenous colloids, iodothyronine lots of colloid, active thyroid when stored. Thyroid gland is lined with single layer of (=thyroxine) follicular cells large, columnar, unique in storing lots of hormone Thyroid lobulated cuboidal follicular cells basal nucleus, less colloid when inactive! scattered, lumps or single cells, endoneurocrine, derived from parafollicular cells calcitonin near fenestrated capillaries for neural crest cells? hormones to enter blood stream poorly defined lobules large, round nuclei, resting cells have chief/principal cells pale cytoplasm, prominent golgi, rER, + septa contained in PTH (most common) secretory granules. When active the capsule of thyroid glandular elements can be smaller, more rER, stain more strongly gland (septa [blue] = intermixed with adipose cells, in Parathyroid extensions of capsule age becomes infiltrated by containing eosinophilic cytoplasm, numerous lymphocytes neurovascular oxyphil cells (minor unknown mitochondria, larger than structures) component) principal cells glandular epithelium, intimate vascular cords or clumps of cells, sinusoid *50% somatrophs (GH) connections with capillaries, collagen and reticulin Anteroir 20% corcitutrophs (ACTH) hypothalamus. GH, ACTH, LH, FSH, network, chromophobes: pituitary blob 20% lactrotrophs (prolactin) chromophobe and prolactin, thyrotrophin smallest, few granules, (adeno) 5% gonadotrophs (LH, FSH) chromophil cells = chromophils: acidophil or 5% thyrotrophs (thyrotrophin) ~troph cells with lots of basophil. granules* axons: lots of granules, accumulate in distended terminations = Herring bodies, Posterior non-myelinated axons, connects to granules contain hormone cell bodies of axons in pituitary pituicytes (speciallised ADH, oxytocin hypothalamus via stalk precursors generated in cell body, hypothalamus (neuro) glial cells) final hormone generated during transport. Pituicyte EM: few granules
  • 18. 3. anatomical structures: GI tract C.Riedinger
  • 19. 4. identify: blood cells (connective tissue) BLOOD Wheater's: table page 64 1. red blood cells: - total absence of organelles - no nucleus - flattened disc with elevated circumference - reticulocytes (= precursors, <1% of circulating erys) have some residual nuclear material - very young cells some rER and mitochondria 2. white blood cells: (1/1000 blood cells) 5 types, named based on staining properties of granules granulocytes: Neutrophils do not stain in humans 60% eosinophils pick up eosin and therefore stain orange 3% basophils pick up azures and therefore stain blue, rarest cells 1% single multilobed nuclei (polymorphonuclear) originally believed to be polynuclear mononuclear leucocytes: lymphocytes clear cytoplasm, rounded nucleus 34% (agranular) monocytes large, indented curved nucleus 4% non-lobulated nuclei agranolucytes 3. platelets: - small - non-nucleated - round or oval, biconvex - cytoplasm purple stained - granules = 20% of platelet volume - many organelles C.Riedinger
  • 20. 4. identify: blood cells (connective tissue) (2) C.Riedinger
  • 21. 4. identify: type of ganglia (nervous tissue) C.Riedinger how to distinguish ganglia cell bodies are large with smaller supporting cells around it symp + parasymp: contain synapses (stellar ganglion = largest symp ganglion) sensory: just contain cell bodies appearance sensory ganglion: many and larger nuclei of satellite (supporting cells), form neat circle around cell body, even larger cell bodies pseudounipolar neurons! sympathetic ganglion: smaller and more scattered satellites, smaller cell bodies more space between cell bodies as axons and dendrites have to pass through! same basic structure as sensory ganglia parasymp. Ganglion: near target organ! Islands of connective tissue with blood vessels, nerves and ducts nerve cell bodies lie within nerve trunk, are surrounded by support cells, less satellites, smaller cell bodies large cell bodies and axons nerves in longitudinal section zig-zaggy strands with nuclei of schwann cells visible each myelin producing schwann cell covers ca. 1mm of the nerve fibre in-between: nodes of ranvier often stained black with pink connective tissue in-between individual myelinated fibres
  • 22. 4. identify: GI tract - looks more structured than stomach C.Riedin organs: alimentary system (gut) - mucin stains more blue than cyan Wheater's: page 286, 287, for glands 97 How to distinguish different parts of the gut: Are there villi? * Yes Small intestine: Duodenum/Jejunum/Ileum Are there brunner's glands? * Yes Duodenum * No Jejunum/Ileum Are there peyer's patches? * Yes Ileum * No Oesophagus/Stomach/Colon * No Jejunum Are there glands? * Yes Stomach/Colon What do the glands look like? Straight and beautiful Colon Thick very thick layer underneath ducts, less Stomach ordered * No Stratified squamout epithelium? Oesophagus
  • 23. 4. identify: urinary system organs: urinary system (kidneys) C.Riedinger adrenal gland egulfed in dense supporting tissue that extends into gland to support secretory cells zona glomerulosa 5-10% secretes mineralocorticoids (e.g. aldosterone) whorls of cells and capillaries zona fasciculata 75% narrow cords of large cels sinusoid capillaries cortex: rich in sER and lipids "foamy" secretes glucocorticoids (e.g. cortisol) zona reticulosa irregular network of branching cords numerous capillaries of wide diameter smaller cells than other two layers secretes androgenic steroids medulla: chromaffin cells clumps and cords of cells surrounded by fine supporting tissue large nucleus (stains blue) basophilic cytoplasm secretes catecholamines (e.g. (nor)adrenaline) steroid secreting cells: many mitochondria with unusual tubular cristae sER lipid droplets (if secreting cholesterol, in cortex) membrane-bound granules (if secreting catecholamines in medulla, but those are not steroids)