Prelim Lab #1 Flashcards by Colleen Gutierrez

Why are there 3 copies needed for Histopath reports

File lab
Physician
Patient

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Signatories

Request

Result

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Routine Turn-over of results

Surgical Pathology (24 hrs)
Frozen section (5-15 mins)
Autopsy report (w/in 1 week)

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Storage of Specimen

Tissue Blocks (3-10yrs)
Slides (indefinitely keep)

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Includes entering the details of the specimen in a logbook.

Numbering

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Gross description is done by who?

A PATHOLOGIST

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Ideal size for specimen processing:

3x2 cm; 3.5 mm thick

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In our experiments the size of specimen processing is?

2x2 cm; 4 cm thick

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Thickness of the lung specimen

2 cm thick

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Preserving the morphology and chemical integrity of the cells as close as to the original as possible.

Fixation

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First and most critical step in histotechnology

Fixation

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Procedures are adopted to kill, harden and preserve materials for microscopic study

Fixation

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Primary goal of fixation?

Preserve

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Secondary goal of fixation

To harden and protect the tissue from trauma of further handling

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Factors involved in Fixation

Temp
Thickness
Osmolality
pH
Concentration 
Volume
Time and Duration of fixation

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Inc in Temp

Heat= accelerates process of distortion

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The temp is traditionally carried out in a?

Room temperature

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Temp. for EM and Histochemistry

0-4 degree C

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Temp. for Autotechnicon

40 degree C

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It is an automatic tissue processor

Autotechnicon

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Formalin used @ 50 degree C

Rapid fixation of urgent biopsies

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Formalin used @ 100 degree C

Tissues with TB

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Thickness for LM

2 cm2

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Thickness for EM

1-2cm2

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ph used in fixation

6-8

Why is the pH 6-8 used in fixation?

For satisfactory fixation

The osmolality of fixation must be in what form?

Isotonic

Effect of Hypotonic

Swellin

Effect of Hypertonic

Shrinkage

Concentration used in Fixation

10% Formalin

Ideal time for fixation

20-30 mins.

Why does the ideal time is 20-30 mins?

Because it is followed by interruption of blood supply

Volume for fixation should be

20x the volume of the specimens

Effects of Fixatives

Harden soft and friable tissue Make cells resistant to damage and distortion Inhibit bacterial decomposition Increase optical differentiation of cells Acts as mordants or accentuations to hasten staining Reduce the risk of infections

Characteristics of a Good Fixative

Cheap Stable Safe to handle Kill the cell quickly Inhibit bacterial decomposition and autolysis Produce minimal shrinkage of tissue Permit rapid and even penetration of tissues Harden tissues for cutting Isotonic Make cellular components insoluble to hypotonic solutions Permit the subsequent application of many staining procedures

Gas produce by oxidation of methyl alcohol

Formaldehyde

Best fixative for nerve tissue

Formaldehyde

Powerful reducing agent

Formaldehyde

40% by weight of Formaldehyde

Stock solution; tends to harden

10% solution of formaldehyde

Working solution; forms pigment

What are the pigments formed in 10% solution of formaldehyde

ParaFormaldehyde and formic

Removal of Formalin Pigments

Kardasewitsch's Method Lillie's method Picric Acid Method

Components of Picric Acid Method

Saturated picric acid

Components of Kardasewitsch's method

70% ethylalcohol and 28% ammonia h2o

Components of Lillie's Method

Hydrogen, acetone, peroxide, and 28% ammonia H20

Preserves cellular and plasma protein better

Glutaraldehyde

2.5 % solution of Glutaraldehyde is for?

Small fragment and small needle biopsies fix for 2-4 hrs at room temp.

4% solution of Glutaraldehyde is for?

Large tissue not exceeding than 4 mm thickness and fix at 12-24 hrs at room temp

For thin and ultrathin sections for plastic embedding

Paraformaldehyde

White powder; used as 4% at 4 degree C

Paraformaldehyde

For excellent routine paraffin section

Paraformaldehyde

For CNS tissues

10% formol saline

Best general tissue fixative

10% Buffered Neutral Saline

Most common metallic fixatives

Mercuric choloride

Recommended for renal, fibrin and CT

Mercuric Chloride

Used 1-2% aqueous solution

Chromate fixatives

Strong oxidzing agent

Chromate fixatives

What are the oxidzing agents used in Chromate fixative?

Chromic Acid Potassium dichromate Regaud's Orth's fluid for Ricketssiae

Chromate fixative that is used in 4% aqueous solution strong fixative for certain lipids

Potassium dichromate

Used for acid muccopolysaccharide

Lead Fixatives

Used in 4% aqueous solution

Lead Fixatives

Best fixative for glycogen

Picric Acid Fixative

Used in a strong or saturated solution

Picric Acid Fixative

Picric acid fixative is chemically known as?

2,4,6 trinitrophenol

Used at ice-cold temperature

Acetic Acid Fixative

Fixes and precipitates nucleoprotein

Acetic Acid Fixatives

Preservation of water diffusible enzymes

Acetone

Best fixative for fixing brain tissue rabies

Acetone

Used only in enzyme studies

Acetone

Used for rapid denaturing and ppt of proteins by destroying hydrogen and other bonds

Alcohol fixative

Concentration of alcohol fixative is?

70-100%

Why does the concentration of alcohol fixative must be 70-100%?

Because lesser concentration lyse the cells.

Commonly known as osmic acid; pale yellow

Osmium Tetroxide Fixatives

Produces excellent nuclear staining

Osmium Tetroxide Fixative

Recommended for materials from the nervous system and general post mortem materials

10% Formol Saline

Fixation time for 10% Formol Saline

12-24 hrs.

Preservation of post mortem surgical research specimen

10% Buffered Formalin

Fixation time for 10% Buffered Formalin

24 hrs and more

Post mortem materials

Formol Saline Sublimate

Fixation time for Formol Saline Sublimate

3-24 hrs.

For routine post mortem materials

Formol Sublimate

Fixation time for Formol Sublimate

3-24 hrs

Fixative for liver and connective tissue fiber

Zenker's solution

Fixation time for Zenker's solutions

3-24 hrs.

Pituitary tissue and bone marrow tissue

Zenker's Formol

Zenker's Formol is also known as?

Helly's fluid

Fixation time for Helly's fluid

12-24 hrs.

Fixative for embryos

Bouin's Solution

Fixation time for Bouin's Solution

6-24 hrs.

Power of sorting; glycogen

Brasil's

Specific for chromosome

Flemming's Fluid without acetic acid

Nuclear structure

Flemming's Fluid with acetic acid

Fixation time for Flemming's Fluid with Acetic Acid

24-28 hrs.

Recommended for chromosomes, lymph nodes and urgent studies of glycogen

Carnoy's Fluid

Fixation time for Carnoy's Fluid

1/3-3 hrs.

Recommended for Mucopolysaccharides, nuclear protein and chromosome

Newcomer's Fluid

Newcomer's Fluid fixation time isv

2-3 hours.

It contains Glacial Acetic Acid

Nuclear Fixative

Does not contain glacial acetic acid

Cytoplasmic Fixative

pH for Nuclear Fixatimve

4.6 or less

pH used in Cytoplasmic Fixative

Greater than 4.6

Preserve fats

Flemming's Fluid with no acetic acid

Fixation time for Flemming's Fluid with no acetic acid

26-48 hrs

For mitochondria, gogli elements and fats

Champy'a Fluid

For mitochondria and yolk

Regaulds

Myolin Fibers, Rickettsia and other bacteria | Study of early degenerative process and tissue necrosis

Orth's Fluid

Cytoplasmic

Zenker's Formol

Removal of Ca ions/ lime salts

Decalcification

Ideal time for decalcification

24-28 hrs

Ratio of fluid to tissue

20:1

Types of decalcification

Acid Method Chelating Agents Ion Exchange Resin Electrolytic Method

Most commonly used method in decalcification

Acid Method

Most common decalcifying agent

Nitric Acid

Combination of chromic acid, nitric acid and ethyl alcohol

Parenyi's

Most rapid decalcifying agent

Phloroglucin-Nitric Acid

Nitric Acid and Formalin

Formol nitric

Best general decalcifying agent

Formic Acid

For urgent biopsies

HCl

Small spicule of delicate tissues

TCA

Small bone

Chromic Acid

Facilitate removal of calcium salts

Chelating agents

Rapid commercial Chelating agents

Cal-Ex

Combines with calcium and forms soluble non-ionized complexes

EDTA

Common name of EDTA

Versene

Decalcification may be carried out in a short period however the degree cannot be measured by chemical means

Ion-Exchange Resins

Most rapid method

Electrophoresis

Carried out by bending the tissue or pricking the tissue with the needle to determine the consistency

Physical/Mechanical method

Most reliable but expensice

X-ray method

X-ray method is not applicable for tissue that has been fixed using whay?

Mercuric Chloride

Chemical method is carried out by transferring how many mL of decalcifying agent into the tube?

5 mL

Histopath Reports

Surgical Pathology Cytopathogy Autotopys report

Prelim Lab #1 Flashcards

(131 cards)