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Foundation

13/02/15

IntroductiontoHistologyandBlood
RBCsare7micronsacross
LightMicroscopy
Maxresolvingpwr=0.2microns=todistinguishtwothingsasdifferentpoints(i.e.cytoplasmic
components)
ElectronMicroscopy
Tissuesprepareddifftolightcutthinly,Stainedwithheavymetals.
Differenttissuecomponentsandmetalstainscausevaryingelectronscattering
Blackandwhiteimageofhighdetail0.2nanometremaxresolutionpower
Fixation
Removingtissuesfrombodyleadstocelllysosomalbreakdownandbacterialinfection
o Needtofixembedsectionstain
Formalin(formaldehyde)usedforfixationinLM,antiseptic,toughentissue
o Fixativescrosslinkmoleculeslockintoplace
Embedding
Tissuemusthavewaterremovedbyalcohol,andreplacedwithxylene(canmixwithmolten
paraffin).Paraffinusedtostiffentissues
Sectioning
Thincut(515micronsthick)cassettespercolatedinparaffin
Sectionspassedthroughalcohol/xyleneremoveparaffinrehydratedputonglassslides
StainingH&Emostcommon
Haematoxylin
- Blue/purple
- Positivelychargedbindstoacidic/negativecomponents(Basophiliccells)
o i.e.DNAphosphategroupsarenegative,RNA,proteoglycans
Eosin
- Pink/orangebindstopositivelycharged
o Negativechargebindstobasicaminogroupsofproteins
(intra/extracellular)Bindseosinophilic/acidophilictissue
components
Amphophilic
- Plasmacellsmakelotsofproteins>lotsofRNAincytoplasm
stainbothH&Epurplishcytoplasm
Immunohistochemistryantibodiesbindtoproteinsaddingsubstrateforenzymecausescolourchange

Eosin

Pink/orange

Binds to cationic tissue


components (those with a net
positive charge)

Cancerhistologyprofile:bigbluepurplenuclei,nucleoli,detailsinnucleus,speckledchromatin,
cytoplasmispink,redcellsveryredeveninH&E

Binds to positively charged amino


4tissuetypes
groups of proteins, whether they
Connectivetissue
be
intracellular (cytoplasm) or
o Fillspaces,createstructure,metabolic,thermoregulatory,defensive
Epithelia
extracellular
o Coverssurfaces(liningoflumens,GIT,coveringofskin,liningofbloodvessels)
Such tissue components are
described as being acidophilic or

Foundation
o Maybesecretory
Muscle
NeuralTissue

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ConnectiveTissue
Twotypes:
Connectivetissueproper
Supportsparenchyma,bloodvessels,epithelia
Loose,Dense:regularandirregular
Specialisedconnectivetissue:Adipose,Bone,Blood,Cartilage,Haemopoietictissue,lymphatictissue

Red blood cells

Blood
-

5L70kgperson,consistsfluid+cells
Function:
o TransportO2/CO2,nutrients,waste,hormones,heat,cells
o Defence
o Haemostasisstoppingthebloodflow
Plasmawater,protein,salts,lipids,sugar
o Fluidinbloodinequilibriumwithextracellularfluid
Plasmaproteins
o Sourceinliver
o 3groups:coagulatingproteins,albumin(acarrierprotein,osmotic
balanceinblood),globulins
Gartner LP & Hiatt JL Colour Textbook
of Histology 2E WB Saunders
o Proteinsexertcolloidosmoticpressure
0721688063 Fig 10.4

platelets

Kerr JB Atlas of Functional Histology Mosby 0723430721 . Fig 2.6

BloodCells
Haematocrit=rbcvolume/bloodvolumenormally45%
RedBloodCells
- Properties:Stainspink/redinH&E
o Biconcavedisc
o Greatersurfaceareathanspheretomax.gasxchangeinlungs/tissue
o Abletosqueezethrucapillaries(rbc7.5micronsdiam)
o Canlooklikeblobwhenaggregated
Reticulocytes
- Nonucleus/organellesAnaerobicmetabolismhasfinitelifespan(120days)
- ContainhaemoglobintransportO2andCO2
Form in which rbcs are
Reticulocytesformofimmatureredbloodcellsasreleasedintoblood
released into blood
- Appearance:
Immature rbcs, no nucleus but
o Slightlylargerthanrbcs,someorganelles/mitochondria,andRNA,Nonucleus
still some organelles and RNA
o Lessthan1%ofcirculatingrbcs
Slightly less than 1% of
- reticulocytesinhaemorrhageorhaemolysisrbcsreleasedwhenstillreticulocytes
circulating rbcs
o orreticulocyte#canbecausedbydifferentabnormalities
Increased numbers in blood
following haemorrhage or
- NormoblastImmaturerbcstillwithnucleus
Platelets
- Appearance:
o 24micronsdiametertinysize
o Containmanygranules
- Action:

haemolysis
Normoblast: immature
nucleated rbc

H&E

Foundation
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o Releasegrowthfactors,thromboxane(vasoc.agent)clottingfactors,havereceptorsimportant
forclottingandcoagulation
o Activatebloodclottingcascadeimportantinhaemostasis(controlofbleeding)
o Plugsmalldefectsinvesselwalls
- lifespan810days

WhiteBloodCells:GranulocytesandMononuclearleukocytes
Granulocytes

Multilobatednucleus,Prominentcytoplasmictypespecificgranules

Neutrophils
Appearance
o 2or3segmentsofnucleusjoinedtoformnuclei
o Granulatedcytoplasm,noorganellesenergyanaerobic
o Twicesizeofrbcs
Madeinbonemarrow
3typesofgranulescontaining:myeloperoxidase,lysozyme,collagenasecontainproteases
Phagocytic,andcapableofdegranulationacuteinflammationrarelyintissuesnormally
Lifetime:severaldays
Eosinophils
Appearance
o Bilobednucleus
o Large,eosinophilspecificgranules
o Verypinkstain
Allergic/parasitereactions
Exocytosesgranules
Basophils
Appearance:
o Bilobednucleus
o Largebasophilicspecificgranulescontainhistaminelikemastcellsintissuevasod.
Degranulateininflamm.Responses
MononuclearLeukocytes
Lymphocytes
B,T,NKcells
Mostlyblue/purplenotmuchcytoplasmmostlydarkstained
o Round,mostlydarknucleus
Monocytes
Appearance
o Largestrbc,oval/beanshapenucleus
o Palernucleusandmorecytoplasmthanleukocyte
Becomemacrophageintissue
o Macophagesstandbyintissuewaitfordamage/infection
o Overallbeanshape,largerthanneutrophil
o Pinkcytoplasm
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Foundation
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Haemopoiesisproductionofrbcs
Inadults:femur,axialskeleton,pelvis
Timelineofhaemopoiesis
o Marrowhemopoiesisoccursinyolksacduringembryo
o Infoetusoccursinliver
o Afterbirthmostlyinbonemarrow
o Inadults,onlyaxialskeletonandprox.Femurcontainredmarrow,moreyellowmarrow
Billionsofcellseachday
AllfromHSCprogenitorselfrenewalfewcirculateinbloodgiverisetoosteoclast
Erythropoiesis
Earliestprogenitorisproerythroblast
Blue/BasophiliclotsofRNA
Maturation:
o Cellsdividegetsmaller
o Formsmallernormoblasts
o Hemoglobinleadstomorepinkstaining
o LatenormoblastexudesnucleustobecomereticulocytewithsomeRNAleft
o Thengoesoffintothebonemarrow
Granulocyteprecursors
Neutrophildevelopment
MyeloblastsandMyelocytesiftheyarepartofthegranulocyteline.
Bigcellthatdivides,developsmoregranules,asmatures,nucleusdivides,endupwithfinalmature
neutrophilwithsegmentednucleus
Matureneutrophilissmaller,goesintoblood
Haemopoiesis
Tightlycontrolledbygrowthfactors(GMCSF)
Micrenvironment
o Cells(mesenchymalSCs,macrophages,fibroblasts)and
o ECM(collagen,reticulin,glycoprotein)
Rapidresponsetohighdemandforextracellse.g.rbcs,neutrophils
Bloodexamination
FullBloodExamination
Rbcshape,number,haemoglobincount
Conc.Ofwbcsandplatelets
Bonemarrowbiopsy
o Bloodfilms,bonemarrowstainedwithRomanovskyWrightsorGiemsastain
Haemostasistests>APTT,INR
Electrolytes,proteins,enzymes,glucose,ironinblood
Focusquestions
1. Describetheprocessoffixing,embedding,sectioning,stainingcellsforhistology
2. Listeachofthemaincomponents(andtheirfunctions)ofcirculatingblood
3. Describethecharacteristicsofeachofthemajorcelltypesfoundinblood
a. Redbloodcells,
4. Describetheprocessofhaematopoiesis
5. Describetheprocessoferythropoiesis
6. Describethedifferentwhitebloodcellsandtheircharacteristics
7. Whatisthekeyinformationobtainedinabloodexamination(FBE),andwhatcantheybeusedfor?
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