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MTAP 2 SPECIFICS > Fixatives HPCT > Flashcards

Fixatives HPCT Flashcards

1
Q

Microanatomical fixatives example

A

10% Formol Saline
10% Neutral buffer formalin
Heidenhein’s susa
Zenker’s solution
Zenker’s Formol (Helly’s solution)
Bouin’s solution
Brasil Solution

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2
Q

Preserver CYTOPLASMIC structure
No glacial acetic acid
pH is more than 4.6

A

Cytoplasmic fixatives

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3
Q

Nuclear fixatives example

A

Flemming’s fluid
Carnoy’s Fluid
Bouin’s Fluid
Newcomer’s Fluid
Heidenhain susa

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4
Q

Cytoplasmic fixatives example

A

Flemming’s fluid without acetic acid
Helly’s Fluid
Regaud’s Fluid (Muller’s fluid)
Orth’s Fluid

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5
Q

Histochemical Fixatives example

A

Formol saline 10%
Absolute Ethyl Alcohol
Acetone
Newcomer’s Fluid

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6
Q

Fixatives for satisfactory for routine paraffin sections
For electron microscopy
For Histochemical and enzyme studies

A

Aldehyde Fixatives

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7
Q

Most widely used concentration for this fixative is 10%

A gas produced by the oxidation of METHYL ALCOHOL

Pure stock solution of this fixative is 40% which is unsatisfactory for routine fixation

Dilution is 1:10 or 1:20

usual fixation time of this fixative is 24 hours

Buffered to pH 7 with PHOSPHATE BUFFER

A

Formaldehyde

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8
Q

Cheap, Readily available, easy to prepare, Relatively stable

Compatible with most stain

Preservers fats, glycogen, and mucin

Allows tissue enzymes to be studied because it does not precipitate proteins

Recommended for nervous tissue preservation

Allows natural tissue colors to be restored; recommended for colored tissue photography

Tolerant fixative used for mailing specimen

A

Advantages of formaldehyde

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9
Q

Disadvantages of formaldehyde

A

May cause sinusitis, allergic rhinitis, excessive lacrimation or allergic dermatitis

May produce considerable shrinkage of tissues

A soft fixative and does not harden some cytoplasmic structures adequately enough for paraffin embedding

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10
Q

Advantages of formalin

A

Cheap, Readily available, easy to prepare, Relatively stable

Compatible with most stain

Preservers fats, glycogen, and mucin

Allows tissue enzymes to be studied because it does not precipitate proteins

Recommended for nervous tissue preservation

Allows natural tissue colors to be restored; recommended for colored tissue photography

Tolerant fixative used for mailing specimen

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11
Q

May cause sinusitis, allergic rhinitis, excessive lacrimation or allergic dermatitis

May produce considerable shrinkage of tissues

A soft fixative and does not harden some cytoplasmic structures adequately enough for paraffin embedding

A

Disadvantages of formalin

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12
Q

Microanatomical fixative

Recommended for fixation of CNS and general postmortem tissues for histochemical explanation

Preserves enzymes and nucleoproteins

Demonstrates fats and mucin

A

10% Formol saline

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13
Q

Recommended for preservation and storage of SURGICAL, POST-MORTEM, and RESEARCH specimen

Fixation time is 4-24 hours

Best fixative for tissues containing iron pigments and for elastic fibers

A

10% Neutral buffered formalin or Phosphate-buffer formalin

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14
Q

Recommended for routine POST-MORTEM TISSUES

Fixation time of this fixative is 3-24 hours

Penetrates SMALL PIECES of TISSUES RAPIDLY

Excellent for many staining procedures including SILVER RETICULUM METHODS

A

Formol-Corrosive or Formol-Sublimate

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15
Q

Fixation of this fixative is FASTER

for RAPID DIAGNOSIS because it FIXES AND DEHYDRATES at the same time

Good for preservation of GLYCOGEN and for MICRO-INCINERATION technique

Used to fix SPUTUM since it COAGULATES mucus

Produces GROSS HARDENING of TISSUES

Causes partial LYSIS of RBCs

Preservation of iron-containing pigments is POOR

A

Alcoholic formalin or Gendre’s fixative

contains 95% ethanol saturated with picric acid, Formaldehyde, and Glacial acetic acid

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16
Q

Made up of 2 formaldehyde residues, linked by 3 carbon chains

For ROUTINE LIGHT MISCROCOPIC WORK

Buffered glutaraldehyde, followed by secondary fixation in osmium tetroxide is satisfactory for ELECTRON MICROSCOPY

Fixation time of this fixative is 1/2 hour to 2 hours

Preserves PLASMA PROTEINS

Produces LESS TISSUE SHRINKAGE

EXPENSIVE

LESS STABLE

Penetrates tissue SLOWLY

Tends to make tissue more BRITTLE

Reduces PAS (Periodic acid–Schiff) positivity of reactive mucin

A

Glutaraldehyde

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17
Q

List of aldehyde fixatives

A

Formaldehyde (Formalin)
10% Formol Saline
10% NBF or Phosphate-buffered formalin
Formol- corrosive / Formol sublimate
Alcoholic formalin / Gendre’s Fixative
Glutaraldehyde

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18
Q

Most common metallic fixative; used in 5-7%
Penetrates poorly and produces shrinkage of tissues
May form BLACK PRECIPITATES of MERCURY
Precipitates ALL PROTEIN
Recommended for RENAL TISSUES, FIBRIN, CONNECTIVE TISSUES, and MUSCLES
Rapidly HARDENS the OUTER LAYER of the TISSUE with incomplete fixation of the center

Trichrome staining is excellent. Permits brilliant metachromic staining of cells

A

Mercuric Chloride

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19
Q

Mercuric chloride stock solution + GLACIAL ACETIC ACID
Recommended for fixing small pieces of LIVER, SPLEEN, CONNECTIVE TISSUE FIBERS, and NUCLEI
Fixation time is 12 - 24 hours

RECOMMENDED FOR TRICHROME STAINING
Permits BRILLIANT STAINING of NUCLEAR and CONNECTIVE TISSUE FIBERS
COMPATIBLE with MOST stains
May ACT as a MORDANT
PENETRATION is POOR

A

Zenker’s Fluid

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20
Q

Fixation time of this fixative is 12-24 hours
EXCELLENT MICROANATOMIC FIXATIVE for PITUITARY GLAND, BONE MARROW, and BLOOD containing organs such as SPLEEN, and LIVER
PRESERVES CYTOPLASMIC GRANULES well

A

Zenker-Formol / Helly’s solution

PBB (Pituitary gland, Bone marrow, BLOOD containing organ)

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21
Q

Recommended mainly for TUMOR BIOPSIES especially of the skin
Excellent CYTOLOGIC FIXATIVE
Fixation time : 3-12 hrs
Produces brilliant results with SHARP NUCLEAR and CYTOPLASMIC details
Permits EASIER sectioning of large blocks
of FIBROUS CONNECTIVE TISSUES
RBC preservation is POOR
Some CYTOPLASMIC granules are DISSOLVED
Weigert’s method of staining elastic fibers is not possible in Susa-fixed tissues

A

Heidenhain’s Susa Solution

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22
Q

commonly used for BONE MARROW BIOPSIES
Rapid fixation can be achiever in 1 1/2 - 2 hours

A

B-5 Fixative

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23
Q

Use in 1-2% aqueous solution
Precipitates ALL PROTEINS and ADEQUATELY PRESERVES CARBOHYDRATES
A STRONG OXIDIZING AGENT
Not used because IT IS HAZARDOUS

A

Chromic Acid

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24
Q

Used in 3% Aqueous solution
PRESERVES LIPIDS AND MITOCHONDRIA

A

Potassium Dichromate

25
Q

Fixation time of this fixative is 12-48 hours
HARDENS TISSUES BETTER and MORE RAPIDLY than Orth’s Fluid
Recommended for DEMONSTRATION OF CHROMATIN, MITOCHONDRIA, MITOTIC FIGURES, GOLGI BODIES, RBC, AND COLLOID-CONTAINING TISSUES
Must always be FRESHLY PREPARED
GLYCOGEN penetration is POOR
NUCLEAR STAINING is POOR
DOES NOT preserve FATS
Intensity of PAS reaction is REDUCED

A

Regaud’s Fluid/ Muller’s Fluid

Preserves GMRC
Golgi bodies, Mitochondria and mitotic fluid, RBC, Colloid-containing tissue

26
Q

Fixation time is 36-72 hours
RECOMMENDED for STUDY of EARLY DEGENERATIVE PROCESSES AND TISSUE NECROSIS
Demonstrates RICKETTSIAE and OTHER BACTERIA
Preserves MYELIN better than BUFFERED FORMALIN

A

Orth’s Fluid

27
Q

Used in 4% aqueous solution of basic lead acetate
Recommended for ACID MUCOPLYSACCHARIDES
Fixes CONNECTIVE TISSUE MUCIN
Takes up CARBON DIOXIDE to FORM INSOLUBLE CARBONATE especially on PROLONGED STANDING

A

Lead Fixatives

28
Q

Normally used in strong saturated aqueous solution (1%)
Excellent Fixative for GLYCOGEN DEMONSTRATION
Also DYES the tissue. ALLOWS Brilliant staining with the TRICHROME method
Precipitates ALL proteins
STABLE
causes RBC HEMOLYSIS and REDUCES the amount of DEMONSTRABLE FERRIC IRON in TISSUES
Must NEVER be washed in water before dehydration
HIGHLY EXPLOSIVE when DRY
ALTERS AND DISSOLVES LIPIDS
SUITABLE for ANILINE Stains
Causes shrinkage of tissue (Slightly Hypertonic)

A

Picric Acid

29
Q

recommended for FIXATION of EMBRYOS and PITUITARY BIOPSIES
Excellent Fixative for preserving SOFT and DELICATE structures
Fixation time of this fixative is 6-24 hours
PRESERVES Glycogen
Does NOT need washing out

A

Bouin’s Solution

30
Q

BETTER and LESS MESSY than Bouin’s Solution
EXCELLENT FIXATIVE for GLYCOGEN

A

Brasil’s Alcoholic Picroformol Fixative

31
Q

Solidifies at 17C
FIXES and PRECIPITATES NUCLEOPROTEINS
Precipitates CHROMOSOMES and CHROMATIN materials
Causes tissue to SWELL (Hypotonic)

A

Glacial Acetic Acid

32
Q

Must be used in concentrations ranging from 70-100% because less concentrated solution will produce lysis of cells

A

Alcohol fixatives

33
Q

Used to fix and preserve GLYCOGEN PIGMENTS, BLOOD, TISSUE FILMS, and SMEARS
Ideal for SMALL TISSUE FRAGMENTS
Excellent for GLYCOGEN PRESERVATION
Preserves NUCLEAR STAINS

Lower concentrations will cause RBC HEMOLYSIS and INADEQUATELY preserve leukocytes
DISSOLVES fats and Lipids

A

Absolute alcohol

34
Q

Excellent for fixing DRY and WET smears, BLOOD SMEARS, and BONE MARROW TISSUES
FIXES and DEHYDRATES at the same time
Penetration is SLOW
Tissues may be OVERHARDENED and DIFFICULT to cut if left for more than 48 HOURS

A

Methyl Alcohol

35
Q

Used for fixing TOUCH PREPARATIONS

A

95% Isopropyl alcohol

36
Q

Used at 70-100% concentration
a SIMPLE FIXATIVE
Fixation time is 18-24 hours
Preserves but DOES NOT fix glycogen

A

Ethyl Alcohol

37
Q

Used to fix BRAIN TISSUES for the diagnosis of RABIES
Fixation time is 1-3 hours
Considered as the MOST RAPID FIXATIVE
Fixes and dehydrates at the SAME TIME
Preserves NISSL’s granules and Cytoplasmic granules WELL
Preserves NUCLEOPROTEINS and NUCLEIC acids
Excellent fixative for GLYCOGEN

A

Carnoy’s Fluid

CURLS -Chromosomes, Urgent Biopsies, Rabies (Brain), lymph nodes/Lymph glands

38
Q

Histochemical fixative and nuclear fixative
Produces BETTER reaction in FEULGEN STAIN than Carnoy’s Fluid
Recommended for fixing MUCOPOLYSACCHARIDES and NUCLEAR PROTEINS
Fixation time is 12-18 hours at 3c

A

Newcomer’s Fluid

39
Q

Most common chrome-osmium acetic acid fixative
Fixation time is 24- 48 hours
Excellent fixative for NUCLEAR STRUCTURES
PERMANENTLY fixes FATS

A

Flemming’s Solution

40
Q

Made up of only chromatic acid and osmic acid
Recommended for cytoplasmic structures particularly the mitochondria
Fixation time is 24-48 hours

A

Flemming’s solution w/o acetic acid

41
Q

Precipitates proteins
WEAK decalcifying agent
Softening effect on DENSE FIBROUS TISSUES facilitates preparation of such sections
POOR penetrating agent
Suitable only for SMALL PIECES OF TISSUES or BONES

A

Trichloroacetic acid

42
Q

Used at ice cold temperature ranging from -5c to 4c
Recommended for study of WATER DIFFUSIBLE ENZYMES especially PHOSPHATES and LIPASES
Used in fixing brain tissues for diagnosis of RABIES
Used as solvent for certain METALLIC SALTS to be used in FREEZE SUBSTITUTION techniques for tissue blocks
Evaporates RAPIDLY

A

Acetone

43
Q

Involves thermal coagulation of tissue protein for rapid diagnosis

A

HEAT FIXATION

44
Q

A process of placing an already fixed tissue in a second fixative

A

SECONDARY FIXATION

45
Q

Form of secondary fixation
2.5-3%K dichromate for 24 hrs to act as mordant for better staining and aid in cytologic preservation of tissues

A

Post-Chromatization

46
Q

The process of removing excess fixative from the tissue after fixation

A

Washing out

47
Q

Solution used for washing out Helly’s solution, Zenker’s Solution, Flemming’s solution, Formalin, Osmic acid

A

Tap Water

48
Q

Solution used for washing Picric’s acid (Bouin’s Solution)

A

50-70% Alcohol

49
Q

Solution used for washing out Mercuric fixation

A

Alcoholic Iodine

50
Q

Fixative of choice and Fixative to avoid when your target to study is PROTEIN

A

Fixative of choice: NBF, Paraformaldehyde
Fixative to avoid: Osmium Tetroxide

51
Q

Fixative of choice and Fixative to avoid when your target to study is Enzymes

A

Fixative of choice: Frozen section
Fixative to avoid: Chemical Fixatives

52
Q

Fixative of choice and Fixative to avoid when your target to study is Lipids

A

Fixative of choice: Frozen section, Glutaraldehyde, Osmium tetroxide
Fixative to avoid: Alcoholic and NBF

53
Q

Fixative of choice and Fixative to avoid when your target to study is Nucleic acid

A

Fixative of choice: alcoholic fixatives
Fixative to avoid: Aldehydes

54
Q

Fixative of choice and Fixative to avoid when your target to study is Mucopolysaccharides

A

Fixative of choice: Frozen section
Fixative to avoid: Chemical

55
Q

Fixative of choice and Fixative to avoid when your target to study is Biogenic amines

A

Fixative of choice: Bouin’s solution and NBF

56
Q

Fixative of choice and Fixative to avoid when your target to study is Glycogen

A

Fixative of choice: Alcoholic fixatives
Fixative to avoid: Osmium tetroxide

57
Q

Fixatives for Electron Microscope (GOPKZ)

A

Glutaraldehyde
Osmium tetroxide
Paraformaldehyde
Karnovsky’s Fixative
Zamboni’s Fixative

58
Q

Used in ELECTRON MICROSCOPY
Preserves CYTOPLASMIC STRUCTURES well such as GOLGI BODIES and MITOCHONDRIA
Produces BRILLIANT NUCLEAR STAINING with SAFRANIN
Adequately fixes materials for ULTRATHIN sectioning in EM
VERY EXPENSIVE
POOR penetrating agent, suitably ONLY for SMALL PIECES of tissues
INHIBITS hematoxylin and makes counterstaining DIFFICULT
EXTREMELY VOLATILE
Can IRRITATE the EYES producing conjunctivitis or may cause deposition of BLACK OSMIC OXIDE in the cornea leading to blindness

A

Osmium Tetroxide

MTAP 2 SPECIFICS (6 decks)

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