Histotechnology

Question

What would not be seen in autolytic tissue?

Answer 1

Nuclear detail lost

Answer 2

Stainability lost

Answer 3

Stain precipitation

Answer 4

Cells coming away from basement membrane

Answer 5

Are 3 cm deep

Answer 6

Must be labeled

Answer 7

Colours indicate day of week the tissue was processed

Answer 8

Has holes to allow the tissue to flow in an out

Answer 9

Functions by creating hydrogen bridges to prevent degredation

Answer 10

Typically shrinks, swells, and distorts tissue

Answer 11

Keeps bacteria and fungi viable

Answer 12

Allows the tissue to be stainable

Answer 13

Density of tissue

Answer 14

Temperature

Answer 15

Type of container

Answer 16

Picric Acid

Answer 17

Acetic Acid

Answer 18

Osmium Tetroxide

Answer 19

Acetic Acid

Answer 20

Gluteraldehyde

Answer 21

It is a coagulant fixative

Answer 22

It is composed of liquid formaldehyde in water

Answer 23

Add methanol to stop the formation of paraformaldehyde

Answer 24

Penetrates tissue slowly but fixes quickly

Answer 25

Picric Acid

Answer 26

Gluteraldehyde

Answer 27

Is a non-coagulant, additive fixative

Answer 28

Can be used to disinfect the cryostat

Answer 29

Can cause false positive reactions in period acid schiff

Answer 30

Creates methylene bridges in tissue to preserve it

Answer 31

Is a component in B5 fixative

Answer 32

Is a common fixative used in the histology lab

Answer 33

Mercuric pigment can be removed using alcoholic picric acid

Answer 34

Is a great fixative when tissue needs to be X-rayed

Answer 35

Fixes lipids

Answer 36

Used mostly for electron microscopy

Answer 37

Commonly a secondary fixative

Answer 38

Is a non-additive fixative

Answer 39

Is composed of picric acid, formaldehyde and mercuric chloride

Answer 40

Is the best preservative for glycogen

Answer 41

Does not stain tissue

Answer 42

Is commonly used as a secondary fixative

Answer 43

Gluteraldehyde

Answer 44

PAF / Zamboni

Answer 45

Picric Acid

Answer 46

Mercuric Chloride

Answer 47

Acetone should not be used for IHC since it destroys enzymes

Answer 48

Acetone is a good fixative since it does not shrink or harden tissue

Answer 49

Ethanol should not be used to fix frozen sections since it inhibits freezing

Answer 50

Ethanol should not be used to preserve glycogen as it is known to dissolve it

Answer 51

Picric Acid

Answer 52

Acetic Acid

Answer 53

Tissue should be placed in fixative 10X greater than the tissue itself

Answer 54

Tissue should have been placed in fixative right after removal from the body

Answer 55

Tissue in fixative should be placed at 4 degrees Celcius

Answer 56

Tissue should have been frozen before fixation

Answer 57

Large tissue should not be fixed

Answer 58

Large tissue must be cut into smaller pieces

Answer 59

Holes must be poked in the specimen to allow fixative to penetrate

Answer 60

Fixative must be injected into the specimen

Answer 61

Uses vacuum technology to infiltrate tissues

Answer 62

Hematoxylin is added to tint the tissue pink for easier embedding orientation

Answer 63

This processor is safer since it does not use heat

Answer 64

This is an open system

Answer 65

The dehydration process begins with the highest alcohol concentration then descends

Answer 66

Ethyl alcohol is the best for dehydration

Answer 67

Butanol is a good substitution because it is quick

Answer 68

Alcohol dehydrates tissue by removing xylene

Answer 69

Tissue would smell like xylene

Answer 70

Tissue would be soft

Answer 71

Impossible to tell until staining

Answer 72

Tissue would be dessicated

Answer 73

First xylene, then several changes of alcohol, then xylene, then wax

Answer 74

Impossible to fix, request new specimen

Answer 75

First several changes of alcohol, then xylene, then wax

Answer 76

Several changes of wax

Answer 77

Improper fixation

Answer 78

Improper dehydration

Answer 79

Improper clearing

Answer 80

Improper infiltration

Answer 81

Chloridine

Answer 82

An example is Limonene

Answer 83

A solution that dehydrates and clears

Answer 84

An example is Xylene

Answer 85

A solution that dehydrates and infiltrates

Answer 86

Increase the temperature of the wax

Answer 87

Remove wax with xylene, dehydrate with alcohol, then clear with xylene, then several changes of wax

Answer 88

Re-fix the tissue

Answer 89

Several changes of wax

Answer 90

Soft wax allows for easy ribboning

Answer 91

Soft wax provides the most support

Answer 92

Hard wax melts at 45 degrees Celsius

Answer 93

Hard wax should not be used if thinner sections are required

Answer 94

Wax should be kept at least 10 degrees over the melting point

Answer 95

Typical melting point is 35-48 degrees

Answer 96

Overexposure to wax will cause tissue hardening and shrinking

Answer 97

Water-bath should be kept 2 degrees above the melting point

Answer 98

Clearing step can be skipped

Answer 99

Infiltration step can be skipped

Answer 100

Dehydration step can be skipped

Answer 101

Microwaves would never be used for processing

Answer 102

Several small biopsies are placed together

Answer 103

Red dot is facing towards you when cutting

Answer 104

Two pieces of small bowel are orientated so that the lumens are facing eachother

Answer 105

Transverse cut of tubular tissue

Answer 106

Incorrect orientation of tissue when embedding - discard the tissue and request new specimen

Answer 107

Poor processing of tissues - check solutions and machine

Answer 108

Tissue is over-dehydrated - increase time in alcohol

Answer 109

NBF precipitate is found in the tubing of the processor - perform a picric acid flush

Answer 110

Block not chilled enough

Answer 111

Cutting too slowly

Answer 112

Dull blade

Answer 113

Block and knife not parallel to eachother

Answer 114

Tissue must be > 3 mm in order to be decalcified properly

Answer 115

Calcium is soluble at pH 4.5

Answer 116

Acid methods are usually slow

Answer 117

Decalcification increases stainability of bone tissue

Answer 118

Mechanical methods such as poking

Answer 119

Chemical methods such as ammonium hydroxide and ammonium oxalate which check for the precipitation of calcium oxalate

Answer 120

Cutting the bone tissue

Answer 121

Radiography

Answer 122

Bone was stained too long in hematoxylin

Answer 123

Bone was underdecalcified

Answer 124

Bone was improperly fixed

Answer 125

Bone was not stained with the correct stain

Answer 126

Disinfect daily with sodium hypochlorite

Answer 127

Use cryostat when rapid diagnosis is required or when fat or enzymes need to be demonstrated.

Answer 128

If a tissue with known TB requires cutting, disinfect afterwards with formaldehyde vapor

Answer 129

If tissue is sticking the antiroll plate may be too warm

Answer 130

Fluorescent microscope for Acid Fast Bacilli

Answer 131

Polarizing microscope for Oil Red O

Answer 132

Compound light microscope for Acridine Orange

Answer 133

Electron microscope for H&E

Answer 134

"Roughing in" is usually done at 20 micrometers

Answer 135

The clearance angle is usually set at 10 degrees

Answer 136

Speed of cutting has no effect on ribbon quality

Answer 137

In a rotary microtome, the knife moves side to side

Answer 138

With linear stainers, time in solution can be varied by adding more containers

Answer 139

With robotic stainers, the programs cannot be changed

Answer 140

Robotic stainers are good for progressive staining

Answer 141

Linear stainers are good for regressing staining

Answer 142

Water bath should be kept at 5 - 10 degrees above the melting point of wax

Answer 143

Tap water should always be used to fill water baths

Answer 144

Parched earth artifact can result if the water bath is too cold

Answer 145

Water bath should be cleaned after each ribbon

Answer 146

Slides for IHC were placed on a hotplate for drying.

Answer 147

Slides with biospy were cut using special techniques with 6 tissues per slide

Answer 148

Slides for neurology were placed in a 37 degree oven for drying

Answer 149

Slides for routine H&E were placed in a 60 degree oven for drying

Answer 150

Frozen sectioning

Answer 151

Placing used blades in the garbage

Answer 152

Fixing tissue that is known to be TB positive with formalin

Answer 153

Storing glacial acetic acid on the counter top

Answer 154

Fixing CJD positive tissue with formalin

Answer 155

metachrome

Answer 156

benzene ring

Answer 157

auxochrome

Answer 158

chromaphore

Answer 159

A method of nuclear staining is acidic dye

Answer 160

A method of nuclear staining is dye + metal mordant

Answer 161

Cytoplasmic staining occurs from the interaction of dye with neutral molecules

Answer 162

Lipids are stained because the dye has a greater affinity to the solution than the fat

Answer 163

Basic dyes will have a negative auxochrome

Answer 164

Basic dyes will produce chloride salt

Answer 165

Acid dyes are cationic

Answer 166

Acid dyes produce magnesium salt

Answer 167

Modifiers will affect staining time

Answer 168

Sulphonics makes acidic dyes basic

Answer 169

Dye binding can be affected by pH

Answer 170

Orthochromic dye will stain a different colour than the dye colour

Answer 171

Prevent mold growth

Answer 172

Enhance staining

Answer 173

Makes the dye more acidic

Answer 174

Makes the dye fluorescent

Answer 175

Hematoxylin

Answer 176

Acridine Orange

Answer 177

Weak acid differentiating basic dyes ex. PTA/PMA for Biebrich Scarlet

Answer 178

Weak base differentiating acid dyes ex. PTA/PMA for Biebrich Scarlet

Answer 179

Excess mordant differentiating ex. iron alum for Verhoeff

Answer 180

Alcoholic differentiation ex. alcohol for eosin

Answer 181

Oxidize the hematoxylin with sodium iodate

Answer 182

Prepare hematoxylin using aluminum since it can withstand strong acids in the stain

Answer 183

After preparing the hematoxylin, pour directly into the coplin jar for staining

Answer 184

Gill's hematoxylin should be prepared for this stain

Answer 185

Gill's hematoxylin for use in Alcian Blue

Answer 186

Mayer's hematoxylin for Oil Red O

Answer 187

Harris hematoxylin for Masson's Trichrome

Answer 188

Weigert's hematoxylin for routine H&E

Answer 189

Hematoxylin stains indirectly

Answer 190

Is never a regressive stain

Answer 191

Cannot be used to stain bone tissue

Answer 192

The best substitute for eosin would be phenol red

Answer 193

Tissue was left in acid alcohol for 20 minutes during coffee break

Answer 194

Aluminum / iron was not present in the hematoxylin

Answer 195

The hematoxylin was not filtered properly

Answer 196

The hematoxylin was stored in a clear glass jar

Answer 197

Tissue was cut at 3 micrometers

Answer 198

Eosin is too concentrated

Answer 199

Eosin was not left in alcohol long enough

Answer 200

Tissue was left in eosin for too long

Answer 201

Places slide in Bouin's for 1 hour at 56 degrees.

Answer 202

Counterstain in Light Green

Answer 203

Places slides in Biebrich Scarlet Acid Fuchsin before PTA/PMA

Answer 204

Uses Mayer's Hematoxylin to stain nuclei

Answer 205

Collagen has stained blue

Answer 206

Cytoplasm has stained red

Answer 207

RBCs have stained red

Answer 208

Nucleic acids have stained red

Answer 209

Ensure than silver solution is neutralized with HCl before disposal

Answer 210

Place the slides in a thin walled coplin jar to ensure good heating in the microwave

Answer 211

Bleach tissue in Oxalic Acid after Potassium Permanganate

Answer 212

Counterstain in Light Green

Answer 213

Gomori Burnter - argentaffin

Answer 214

Grocott - argyrophilic

Answer 215

von Kossa - substitution

Answer 216

Gordon and Sweets - argyrophilic

Answer 217

Elastic fibers cannot easily be seen on the brain control slide so it was placed in Verhoffs stain for a longer period of time

Answer 218

Nuclei are not staining so a new solution of Verhoffs made and used

Answer 219

The stain looks very muddy so it was placed back into gold chloride for a longer time

Answer 220

Elastic fibers are too pale so the slide was placed back into van Gieson stain for longer

Answer 221

Orcein is specific for elastic fibres

Answer 222

Orcein will stain elastic fibres brown

Answer 223

Aldehyde fuchsin will stain elastic fibres green

Answer 224

Aldehyde fuchsin is a regressive method

Answer 225

Take the slides down to water before staining

Answer 226

Place the slides in Toludine Blue to stain for mast cells

Answer 227

Place the slides in Light Green as a counterstain

Answer 228

Differentiate with alcohol then clear and mount

Answer 229

Stomach tissue was used as the control

Answer 230

The tissue was placed in 3% acetic acid before Alcian Blue dye

Answer 231

The tissue was placed in Acetic Acid following Alcian Blue for 10 minutes

Answer 232

The pH of Alcian Blue was 6.5

Answer 233

Schiff's reagent was tested before use using formalin and the reaction became a deep blue colour

Answer 234

Grocott's method was performed earlier so the periodic acid solution was saved and used agian

Answer 235

Metabisulfite rinse was prepared using 100% HCl straight from the bottle

Answer 236

Tissues were washed for 10 minutes in water after Schiff's

Answer 237

Schiff's reagent is composed of Acid Fuchsin and Sulpheric Acid

Answer 238

Periodic acid is the secondary oxidizer

Answer 239

Sodium Metabisulfite rise is what brings the pink colour to the tissue

Answer 240

Fungi will stain pink

Answer 241

Metanil Yellow is the primary stain

Answer 242

Mucicarmine stains acid mucins

Answer 243

Gill's hematoxylin should not be used as the nuclear stain

Answer 244

Cryptococcus can also be demonstrated with this method

Answer 245

The technologist double checked to ensure sections were cut at 4um

Answer 246

The doctor would like to differentiate between primary and secondary amyloidosis so the slide was pretreated with potassium permangonate

Answer 247

Slides were placed in neutral red to stain for amyloid

Answer 248

The technologist did not know how to use the polarizing microscope so birefringence was not checked on the control

Answer 249

The tissue was previously fixed in formalin

Answer 250

A frozen section was cut for staining

Answer 251

Tissue was taken down to water before staining

Answer 252

The tissue was placed in hematoxylin after Oil Red O

Answer 253

Luxol Fast Blue dye is differentiated with an acid

Answer 254

The control tissue is cerebellum

Answer 255

Eosin is the counterstain

Answer 256

Luxol Fast Blue stains myelin

Answer 257

Tissue is placed in silver nitrate and then microwaved for the appropriate time

Answer 258

Placenta is used as the calcium control

Answer 259

The tissues are placed in nuclear fast red or neutral red

Answer 260

The technologist decides the control passed since the calcium stained brown

Answer 261

Slide was placed in eosin

Answer 262

Patient tissue does not contain iron

Answer 263

Slide was taken down to distilled water

Answer 264

Slide was placed in HCl + Potassium Ferrocyanide

Answer 265

This stain is used to demonstrate argentaffin granules

Answer 266

Gold chloride is a fixing agent

Answer 267

Hematoxylin is the counterstain

Answer 268

Melanin cannot oxidize silver therefore a reducer such as Sodium Thiosulfate is required

Answer 269

Ziehl Neelson

Answer 270

After fixation, Gram stain the tissue to detect the bacteria

Answer 271

Wear N-95 for frozen section

Answer 272

Place the specimen in Carnoy's for fixation then stain with Ziehl Neelson

Answer 273

After fixation, take down slides to tap water before staining

Answer 274

Not enough Acid Alcohol was used to decolourize the AFB

Answer 275

Methylene blue stain was missed

Answer 276

Nature of their cell wall

Answer 277

Because they are Gram negative

Answer 278

Crystal violet - stains Gram negative bacteria

Answer 279

Acetone / Alcohol - decololourizes Gram positive bacteria

Answer 280

Basic Fuchsin - stains acid fast bacteria pink

Answer 281

Picric Acid - stains bacteria yellow

Answer 282

Fungi are stained black - the slide is placed back into gold chloride for more toning until fungi are brown

Answer 283

Fungi, nuclei, and other tissues are staining black - the slide is good

Answer 284

No fungi have been stained - the wrong concentration of periodic acid was used

Answer 285

Fungi are brown - the slide is good

Answer 286

MSB stain for bile

Answer 287

Warthin-Starry for H. pylori

Answer 288

Steiner for spirochetes

Answer 289

Hall for fibrin

Answer 290

Deparaffinize and hydrate to water

Answer 291

apply antibody

Answer 292

stain nuclei with hematoxylin

Answer 293

counterstain with hematoxylin

Answer 294

Tissue must be washed with water to remove crosslinking from formalin

Answer 295

Proteolytic enzymes will destroy the antigen sites

Answer 296

Heat methods help cleave the bonds

Answer 297

Enzymatic methods will unfold the bonds

Answer 298

Picric acid in a container

Answer 299

Alcohol in a container

Answer 300

Sodium Thiosulfate down the sink

Answer 301

Silver Nitrate down the sink

Answer 302

Kidney for Masson's Trichrome

Answer 303

Placenta for von Kossa

Answer 304

Cerebrum for Luxol Fast Blue

Answer 305

Stomach for Alcian Blue

Answer 306

To prevent the blade from getting scratched

Answer 307

To prevent your hands from getting infected

Answer 308

To prevent skin cells from getting on the microscope slides

Answer 309

To prevent yourself from getting cut

Answer 310

Left in Sodium Thiosulfate for too long

Answer 311

Left in Ferric Chloride too long

Answer 312

Left in Van Gieson stain too long

Answer 313

Left in Verhoeff stain too long

Answer 314

pH of eosin is 8.0

Answer 315

Sections were cut at 8 micrometers

Answer 316

Tissue left in alcohol for too long

Answer 317

Tissues were rinsed in water after blueing

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