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BVetMed - Complete course (Years 1, 2, 3) > Histology > Flashcards

Histology Flashcards

All information that was taught to me while attending Vanier College's "Animal Health Technology" Program, located in St-Laurent Montreal.

1
Q

What is histology

A

study of the microscopic anatomy of cells and tissues of plants and animals~sectioned, stained and mounted on a microscope slide~enhanced through the use of histological stains

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2
Q

What is histopathology

A

the microscopic study of diseased tissueimportant tool in anatomical pathology

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3
Q

What are the two types of histological samples that can be taken

A

from a dead animal (necropsy)from a live one (biopsy)

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4
Q

What are the components of the histology lab

A

Fixation of tissuesDehydration, clearing and embeddingSection cuttingParaffinCryostatStaining sections and mounting coverslipsPhotomicrography

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5
Q

What is the goal of histological techniques

A

evenly sectioned (no thick and thin areas),nicely stained (not over or under stained) with the appropriate stainno artifacts (rips, tears, air bubbles, bits of dirt or crystals of stain)in a condition to last many years (properly fixed, coverslip properly mounted)

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6
Q

Describe fixation of the tissues

A

To fix the physical state of the cells, as well as the chemical state. Allows for the subsequent treatment of the tissue with minimal damage and alteration of the tissue. Must not interfere with cell components that are active in staining (or they won’t stain)

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7
Q

What are characteristics of a well fixed slide

A

good nuclear and cytoplasmic morphologyminimal shrinkage clearly defined basement membranes and cell margins.

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8
Q

What are the characteristics of a badly fixed slide

A

Inferior nuclear and cytoplasmic morphologyexcessive shrinkage and poorly defined cell margins

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9
Q

What is very important for the technician for fixation of cells

A

Kills microorganisms~prevents tissue deterioration~protects the technician handling the tissue from pathogens which might be present.

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10
Q

What are examples of chemical fixatives

A

Formalin: most commonSafe-fix: less toxicOther:Special for electron microscopyCryostat

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11
Q

During fixation can tissue deteriorate?

A

Yes.Tissue can deteriorate very rapidlyE.g. Bone marrowMetabolically active deteriorate fastestKidney, liver, pancreasShould be removed first

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12
Q

What is an alternative form of tissue fixation

A

An alternative method of tissue preservation is perfusion of the animal (often used in research). Removal of blood from organs by perfusing with physiological salineImmediate entry of the fixative (normally 10% formalin) into all the blood vessels

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13
Q

What are the necessary components of dehydration, clearing and embedding

A

Gets the tissue ready for sectioning (except for sectioning in a cryostat)We are embedding the tissues in waxMust remove water from the tissues (replacing with a fluid soluble in both water and wax) (dehydration and clearing). Cannot be done all at once (would damage the tissue), so series of steps. Each step is at least one hour, so the process is automated.

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14
Q

How do you dehydrate tissue sample

A

Remove waterUsing alcohol in increasing concentrationMakes the tissue firm for cuttingPrevents shrinkage in paraffin

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15
Q

How do you clean the tissue sample

A

Replaces the alcohol with a liquid compatible with paraffinIncreases tissue transparencyToluol and xylene are mostly used

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16
Q

How do you embed the tissue sample

A

Makes tissue firmer to prepare for actual block preparationThis last step needs to be timed so that the tissue can be removed as soon as it is ready

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17
Q

Why do we trim the tissues before embedding

A

smaller pieces should be cut for treatment, so that a cut surface suitable for sectioning is prepared. This is also a time when excess fatty and connective tissue can be cleaned off the outside of an organ.

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18
Q

Describe block embedding

A

Done at the embedding machine, which has a liquid paraffin dispenser. set at 56C-60CA thin layer of paraplast (liquid) is put in plastic embedding mold.The paraplast impregnated tissue is then addedThe paraplast should have cooled slightly, just starting to form a thin skin, when the tissue is placed in it.

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19
Q

What is the microtome and what is it used for

A

machine equipped with a very sharp knife and a mechanism to advance the tissue in very small increments. Tissues are normally sliced at 6 - 10 microns. We use a rotary microtome, which uses a wheel to advance the tissue block.The knife should be clean, sharp and cold for best sectioning. Sharpen the knife and store it in the freezer (or overnight) before use.

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20
Q

What are the steps for cutting things with a microtome

A

Fasten block Adjust for straightnessBring forward blade so block is almost or just touching. Cut (advance to tissue) (10-15 microns) until a whole section is being taken. When ready to take good sections, be sure the cutting part of the knife is sharp (reposition it if necessary). Reduce thicknessTurn the wheel slowly and smoothly to get even sections (hard tissues can be cut more quickly). The sections should come off in a ribbon.

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21
Q

How do you float tissues

A

The ribbon is placed on paper or in box, and the sections are cut apart and transferred to a floating-out bath which contains a very weak gelatin solution at about 45C .Alternately a ribbon can be transferred to the bath and then the sections separated. Ribbons need to be spread out.

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22
Q

Describe staining tissues

A

The goal of a stain or stains is to allow examination of the various characteristics and relationships of the cells.Different tissues, and different cell components, attract different dyes and stains. Hematoxylin and eosin, the most commonly used pair of stains, are attracted by different cell components,

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23
Q

Describe H/E staining

A

Slide must be dryStains are in solutionFor paraffin-impregnated tissue sections, the wax is removed by xylene, then the tissue is rehydrated by moving from 100% to 95% to 80 or 70% alcohol. The tissue is then placed in water before staining. After staining the tissue section is dehydrated with 95% and 100% alcohol, and the alcohol is removed by xylene. The slide is now ready to have its coverslip mounted.

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24
Q

what are the 2 main functions of the mounting medium

A

It has a refractive index close to that of glass so that the tissue and the slide match It protects the tissue from physical and chemical injury.

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25
Q

What are some types of artifacts on a slide

A

Air bubblesfold of tissuesknife crackshairdebris

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26
Q

What is a laboratory diagnosting test dependant on

A

Good historyQuality of sampleProper identification of sample

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27
Q

How do you choose what sampling technique to use for histology

A

Anatomic locationPatient’s overall healthSuspected tumor typeClinician’s preference

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28
Q

What are the pretreatment biopsy types

A

Needle core biopsyPunch biopsyWedge biopsy

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29
Q

How do you obtain additional information about a tumor

A

treatment planning (surgical, medical)

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30
Q

What is excisional biopsy

A

Surgical removal of the tumor

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31
Q

what is a post treatment biopsy method

A

excisional biopsy

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32
Q

How do you obtain a more complete picture about a growth

A

GradingLymphatic/vascular invasionMargins

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33
Q

Is a biopsy a good first step?

A

No

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34
Q

What is the disadvantage to not doing a biopsy first when a lump is removed

A

can result in incomplete removal, more morbidity and costs

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35
Q

What are the advantages to pre-treatment biopsies

A

Can help clients make an informed decisionCan consult with oncologist and surgeonCan plan treatment sooner after surgery

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36
Q

When are pre-treatment biopsies not indicated

A

Treatment or Sx would not change (spleen, testicle)As risky as removal (spinal cord)

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37
Q

What are needle core biopsies done on

A

external palpable masses (no highly inflamed or necrotic)deep (kidney, liver)

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38
Q

Describe the needle core biopsy punch

A

manual or spring/pneumatic poweredSmall sample size still enough for pathologic exam

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39
Q

what is the size of the needle core biopsies needle

A

1 mm wide biopsy1.0 – 1.5 cm long

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40
Q

What does a needle core biopsy require

A

local anesthesia and sedationsterile preparation

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41
Q

Why do you use a small scalpel incison for the needle core biopsy

A

Prevents dullingFacilitates tru-cut mechanismCan be sutured

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42
Q

How do you handle the tissue from the needle core biopsy

A

Tissue can be removed with blade, needle or salineCan be rolled on glass slide for cytologyPlace in formalin (in cassette)

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43
Q

What is a possible risk when you do a needle core biopsy

A

minimal risk of seeding but you should plan ahead and remove original incision tractconsider hemorrhage and fluid leakage

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44
Q

Why do you use a punch biopsy

A

Typically for skinSkin, oral, perianalDirect access with laparoscopyLiver, GIT, etc.

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45
Q

What is the size of a punch biopsy

A

2-8mm

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46
Q

What is required for punch biopsy

A

local anesthesia and sedationusually no sterile preparation

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47
Q

what is the ideal size of a punch biopsy

A

6mm4 mm only for nose, footpad8 mm slight more chances of infection

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48
Q

What can cause tissue compression and artifacts when doing a punch biopsy

A

dull punches

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49
Q

how do you handle a tissue sample from a punch biopsy

A

handle sample very gentlyplace in formalin, no cassette

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50
Q

what is important for punch biopsies if you’re doing dermatology

A

draw line in direction of hair

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51
Q

When do you do an incisional biopsy

A

When cytology and/or biopsy is unsuccessfulFor ulcerated and necrotic lesions (larger sample)

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52
Q

What do you need to do for an incisional biopsy

A

Surgical preparation + drapesLocal anesthesiaTumors are usually POORLY innervatedSkin is incised and tumor wedge removed

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53
Q

Is it necessary to remove intact skin with the incisional biopsy?

A

NOT necessary to remove intact skin (next or over)Margins evaluated with removal of tumorCan compromise Careful not to sample just the reactive tissue surrounding the tumorImprint cytology can be done

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54
Q

Describe endoscopic biopsy

A

Convenient, cost-effective, safeLimited sample, inadequate visualization

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55
Q

Describe laparoscopy, thoracoscopy

A

Very good, can always convert to laparotomyNeeds specialized tools & skills

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56
Q

How do you properly identify margins

A

tissue ink is preferred but sutures can also be used.need to write: color = which margin.

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57
Q

when should tissue ink be done

A

before fixation, to orient the sample and identify areas of concern

58
Q

describe the tissue inking process

A

Tissue should be blotted with paper towel beforeInk can be applied with gauze on surface or cotton swab for precisionAllow to dry 20 min before formalin

59
Q

describe proper tissue fixation

A

10% buffered neutral formalinSpecial fixative for eyes, testicles1 part tissue to 10 part fixativeIdeally 1 container per lesiondone within 30 minutes

60
Q

describe containers for fixation

A

Wide container, secure lidNo glassNo more than 1LIn secure plastic bag (ziploc)With absorbent packing materialInsulated (avoid freezing)

61
Q

Descriibe large unusual specimen fixation

A

Amputated limbs, spleenCan overnight on iceCan pre-fix 48-72h with partial parallel incisions approximately 1 cm apart (‘‘bread loafing’’)Then shipped without formalin (double-bag) or in 1:1 formalinCan section and fix/send in individual containersAn annotated digital image or sketch of the original specimen to depict sectioning and orientation should accompany the samples

62
Q

describe very small unusual specimen fixation

A

Labeled cassettesDo not use gauze sponges or cardboard because tissue may become compromised upon retrieval

63
Q

describe luminal organs unusual specimen fixation

A

Flush the intact lumen with formalinPartial longitudinal incision3 labeled sections (cranial/proximal, mass and caudal/distal) can be submitted

64
Q

describe thin flat sample fixation

A

Small: placed in a tissue cassette with a foam pad to minimize tissue curlingLarger samples can be tacked onto a flat piece of cardboard presoaked in formalin or water with suture through edges of tissue not needed for examination

65
Q

What do you need to write about the lesion in the mass submission form

A

Lesion-specific clinical history (eg, anatomic site, date first noticed, rate of growth)Potentially lesion-associated clinical signs (eg, lameness, vomiting)Type of lesion (eg, new lesion, recut following incomplete excision, excisional biopsy following previous incisional biopsy, local recurrence)Results of prior lesion-associated diagnostic tests - cytology, prior biopsy reports, imaging (radiographs, ultrasound, MRI, CT); access to radiographs may be especially important for bone and gingival tumors.

66
Q

What general information do you need to put on the mass submission form

A

General clinical history—previous neoplastic diseases, previous or current nonneoplastic conditions of relevanceTreatment history—local and systemic, current and previous (eg, chemotherapy, radiation, corticosteroids)Previous unrelated treatments or potential tumor-inducing historical events at tumor site (eg, previous radiation, vaccination, implants)

67
Q

What other abnormalities should you include on the mass submission form

A

CBC, biochemical, and hormonal (eg, hyperinsulinemia) abnormalities

68
Q

What else should be on the submission form

A

Working clinical diagnosis and/or list of differentialsThorough gross lesion description.Indication whether the submitted sample is an incisional or excisional biopsy. Excisional biopsy indicates assessment of surgical margins is necessary, whereas for incisional biopsies the margin evaluation is null. Anatomic site should be thoroughly describedFeatures appreciated during diagnostic imaging or perioperatively should be describedtissues involved or associated with the mass (eg, thyroid mass invading subjacent skeletal muscle).

69
Q

What is cytology

A

examination of cells having exfoliated from tissue or having accumulated in body fluid

70
Q

what can cytology provide

A

May provide definitive diagnosisVery dependent on sample quality!

71
Q

is cytology invasive?

A

Not invasive

72
Q

How quickly does cytology need to be processed

A

rapidly

73
Q

What should be in the cytology kit

A

ClippersCleansing & disinfectant wipesSyringes: 6 – 12 ml, up to 20 mlNeedles: 1 – 1.5 in (20g to 22g), 2.5 – 3.5 in spinal needle with styletBone Marrow aspiration needle & core biopsy materialScalpel bladesCulture swabs & applicator sticks for slide preparationBox of slides (frosted)EDTA and red top tubesRigid, flat surface for 6 -10 slides (foam tray)Butterfly catheter, IV extension tubingPencil or slide markerSterile EDTA

74
Q

What is the skin prep for a FNA

A

Minimal for cutaneous, subcutaneousShave, clean & disinfect for internal

75
Q

Where can you do an FNA

A

Cutaneous, subcutaneous, internal organs

76
Q

What are the benefits of going needle only, vs aspiration

A

Should start with no aspirationLess blood contamination

77
Q

What is the best FNA size needle

A

22g

78
Q

What do you do if a FNA sample is liquid

A

place in edta

79
Q

what do you do if an FNA sample is solid

A

prepare slides immediately

80
Q

Describe imaging guided aspiration

A

typically ultrasound guidedcomplications are rarethoracic samples can be taken.

81
Q

Should ascites and pleural effusion be sampled via US

A

yes

82
Q

Describe the squash preparation

A

Most commonFor semi-solid, mucus-like, or pelleted (via centrifugation)Place sample close to frosted edge and use second slide to spread sample

83
Q

What do you do with fluid cytology samples

A

Keep in EDTAPrevents clotting (fibrin)Preserves cellular morphologyFacilitates cell countRefrigeratedUp to 24h in generalCSF needs special measuresMake slide as soon as possibleSend unstained slide with EDTA tube

84
Q

How do you prepare a fluid slide if the sample is cloudy

A

direct smear (like blood smear)squash

85
Q

how do you prepare a fluid slide if the sample is clear

A

If lower cellularityNeed to concentrateSimilar to urine sedimentVia special centrifuge (CSF, BAL)Buffy CoatStill do a direct smear

86
Q

What hematology methods can you use with cytology fluid

A

Cell countsCan be automated (hematology machine)Flush with saline afterManuallyTotal proteinsRefractometerConversion tables for low TP

87
Q

Describe the touch imprint

A

Permits evaluation of a biopsyWith surface lesions is often of poor diagnostic utilitySuperficial inflammation, secondary bacterial infectionException for fungal diseasesNeed to blot aggressively the sampleUntil tackyThen imprint on slideFibrous lesions can be scrapedScalpel blade

88
Q

How do you prepare joint-synovial fluid cytology

A

Surgical preparationNormally viscousIdeally small EDTA tubeTo get cell countTo be able to use hyaluronidazeDirect smear (like seen previously)Culturette for microbiology

89
Q

What are miscellaneous types for doing cytology

A

Always air dry – do not use flameDo not freezeDo not expose to formalinLabel properly all tubes & slidesAlways submit an unstained smear with a tube (also for hematology)

90
Q

When do you do bone marrow evaluation

A

Bone marrow evaluation is indicated when peripheral blood abnormalities are detectedpersistent neutropenia, unexplained thrombocytopenia, poorly regenerative anemiaTo stage neoplasiaLymphoma, plasma cell tumor, mast cell tumors, other

91
Q

What is the difference between aspiration and core biopsy

A

Aspirates are easier, faster, and less expensive to perform than are core biopsies. Bone marrow core biopsies require special needles. Core biopsy sections provide a more accurate way of evaluating marrow cellularity and examining for metastatic neoplasia than do aspirate smears, but cell morphology is more diffcult to assess.

92
Q

How can you classify inflammation

A

purulentpyogranulomatousmacrophagiceosinophiliclymphocytic

93
Q

Describe purulent inflammation

A

predominance of neutrophils (>85%)Try to say if degenerated or not

94
Q

Describe pyogranulomatous inflammation

A

mix of neutrophils and macrophages

95
Q

Describe macrophagic inflammation

A

predominance of macrophages (>50%)

96
Q

Describe eosinophilic inflammation

A

important component of eosinophils (>10 – 30%)

97
Q

describe lymphocytic inflammation

A

need to rule out lymphoma

98
Q

What does it mean if you have degenerate neutrophils in your purulent inflammation

A

Degenerate neutrophils:Bacterial infectionsNeed to see to call septic

99
Q

What are the causes of degenerate neutrophils in purulent inflammation

A

Degenerate neutrophils:Immune-mediatedNeoplasticSterile irritants (bile, urine)

100
Q

What are granulomatous lymphocytes associated with

A

Foreign bodyFungal infectionMycobacterial infectionPanniculitisLick granulomaOther chronic lesions

101
Q

what is eosinophilic inflammation due to

A

Eosinophilic granulomasHypersensitivityParasitesFungalMast cell tumorsSome neoplasms

102
Q

what is lymphocytic inflammation due to

A

RareImmune reactionViralChronic lesion

103
Q

What is the infectious agent blastomycosis associated with

A

Pyogranulomatous or granulomatousDogs mostly“hunting”Nose, legsFound in the environment

104
Q

What is the infectious agent cryptococcus associated with

A

GranulomatousDogs & CatsNose

105
Q

where is the aspergillus fungi found

A

OpportunisticDog: noseHorse: cornea

106
Q

Describe mycobacterium

A

Fairly rareGranulomatousVery slow to grow in microbiology

107
Q

How do you classify a neoplasm

A

need to say the cell typeif it is benign or malignant

108
Q

what are the 4 cell types of neoplasms

A

epithelial cellsmesenchymal cellsround cellsneuroendocrine cells

109
Q

how do you classify if a cell is benign or malignant

A

Set of criteriasCytoplasmicNuclear

110
Q

what are the cellular features of malignancy

A

Cellular Crowding Pleomorphism Different shapesAnisocytosisDifferent cell sizeGiant cellsBasophiliaHigh N/C ratio (nuclear/cytoplasmic)Not always

111
Q

what are the nuclear features of malignancy

A

Nuclear Nuclear moldingMore than 1PleomorphismeAnisocaryosisWithin a cell alsoMitotic figureNumber and shape

112
Q

what are the nucleolus features of malignancy

A

multiplevaried within one nucleusmay be normal

113
Q

Describe the organization of mesenchymal cells

A

Weak cohesion, loosely arrangedOften extra-cellular matrix

114
Q

describe the cellular types of mesenchymal cells

A

Cellular typesEg. fibroblasts, osteoblasts, chondroblasts…

115
Q

describe the morphology of mesenchymal cells

A

Morphology Spindled, stellate, ovalPoorly defined cytoplasmic margins

116
Q

describe the exfoliation for mesenchymal cells

A

ExfoliationModerate to weak

117
Q

describe the organization of epithelial tumors

A

Cohesive clusters

118
Q

describe the cell types of epithelial tumors

A

Glandular and parenchymal tissueSurface liningEg. Basal cells, squamous cells, hepatocytes, tubular cells, renal cells

119
Q

describe the morphology of epithelial tumors

A

Variable: round to polygonal +/- elongatedDistinct cytoplasmic borders

120
Q

describe the exfoliation of epithelial tumors

A

very easy

121
Q

What are all the round cell types

A

LymphomePlasmocytomeMastocytomeHistiocyteSarcome histiocytaire Transmissible veneral tumor

122
Q

What does it mean when you see lymphoglandular bodies

A

Cytoplasm fragmentMostly seen with lymphocyteslymphoma

123
Q

what does it mean when you see collagen breakdown

A

Mostly seen in mast cell tumorsSome soft tissue sarcoma

124
Q

what does it mean when you see a hematoidin crystal

A

Hematoidin crystalHemoglobin breakdownIndicates chronic bleeding

125
Q

what does it mean when you see cholesterol crystals

A

Cholesterol crystalsCell membrane damageFrequent inFollicular cysts

126
Q

what does it mean when you see skeletal muscle

A

Skeletal muscleNormalIncidental finding

127
Q

How do you histologically recognize the liver

A

Solid tissue, characterized by cuboidal cells in linesMay or may not see cortex of thin connective tissueLittle C.T. , so not good for trichrome stains

128
Q

How do you histologically recognize the spleen

A

Solid tissue like liver, H and E, stains darkVarious cell types, notice patches of colors at high power differentiating white and red pulp

129
Q

How do you histologically recognize the brain

A

Solid tissue, look for characterictic grey and white matter, and Gyri (round, curving areas)Cerebellum is also helpfulDifficult to get a good cut, so keep trying

130
Q

How do you histologically recognize the kidney

A

Remember, nephron- look for glomeruliAppear as nest like structures, near cortex (outside)Also easy to ID renal pelvis

131
Q

How do you histologically recognize the intestines

A

In general, all should be cross-sectional, and have a lumenDuodenum, jejenum and ileum all resemble each other, just differentiate by looking for peyers patches, length of villi..etc…

132
Q

How do you histologically identify the colon

A

Slightly different with larger villi, and more prominent muscularis layer for larger peristaltic action

133
Q

How do you histologically identify other tubular structures

A

Differentiate uterus from esophagus/trachea from intestinal structuresUterus has more CT, and large endometrium, myometriumUterus Great for trichrome

134
Q

How do you histologically recognize esophagus/trachea

A

Usually can pick up cartilage cells on trachea, large and clear. Esophagus often collapsesUsually the 2 are together

135
Q

How do you histologically identity lymph nodes

A

Solid structures, with white and red pulp like spleenDifferentiate by shape, and amount of red pulp in circles

136
Q

how do you histologically identify glands

A

All glands resemble each other by having acini filled with somethingDifferentiating salivary versus thyroid versus prostate gland is challenging.

137
Q

How do you histologically identify ovaries and testis

A

Look for characterized cells of eachOvaries have ova at different stages, CL is also easy to see Look for the hilus

138
Q

How do you histologically identify testis

A

Look for seminiferous tubules which take up most of the structure, and will have sperm in them

139
Q

How do you histologically identify the esophagus

A

This is esophagus, longitudinal section. Note the stratified squamous epithelium

140
Q

how do you histologically identify muscle

A

Striated muscle at high power- all look the same, lots of elastic fibers

BVetMed - Complete course (Years 1, 2, 3) (26 decks)

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