Type

Aldehyde Fixatives
satisfactory for routine paraffin sections, for electron microscopy and when histochemical and enzyme studies are indicated.

Name Formaldehyde (formalin)

Formula

Fixation time 24 hours

Notes 10% formalin is one of the most widely used A gas produce by the oxidation of methyl alcohol Usually buffered to pH7 with phosphate buffer Simple microanatomical fixative made up of saturated formalin (40%wt vol) CNS, and gen. post mortem procedures Preservation and storage of surgical, post-mortem and research specimens Best fixative for tissues containing Fe pigments and elastic fibers

10% Formol citrate

Formaldehyde, 40% [100mL] NaCl [9 gm] Distilled water [900 mL] Sodium dihydrogen phosphate (anhydrous) [3.5gm] Disodium hydrogen phosphate (anhydrous) [6.5gm] Formaldehyde 40% [100mL] Distilled water [900mL] Sat. Aq. Mercuric chloride [90mL] Formaldehyde 40% [10mL]

24 hours @35C (95F) 48 hours @20-25C (65-77F) 4-24 hours

10% Neutral Buffered Formalin or Phosphatebuffered Formalin (pH 7)

Formal-Corrosive (FormalSublimate)

3-24 hours

Alcoholic Formalin (Gendres) Fixative

95% ethyl alcohol saturated with picric acid [80mL] Strong formaldehyde solution [15mL] Glacial acetic acid [5mL]

Post-fixation with phenol-formalin for 6 hours or more

Formol mercuric chloride ( routine post mortem) Penetrates small pieces of tissues rapidly No need for washing-out Forms mercuric chloride deposits Inhibits determination o fextent of tissue decalcification Fixation is faster (about ) Rapid diagnosisfixes and dehydrates at the same time Good for preservation of glycogen and for micro-incineration technique Made up of 2 formaldehyde residues linked to 3 carbon chains Buffered gluta+ secondary fixation with osmium tetroxide is satisfactory for Electron microscopy 2.5%--small tissue fragments and small biopsies fixed 2-4 hours at RT 4%--larger tissues <4mm thick, fixed 6-8 hrs up to 24 hrs. Most common metallic fixative Satd aq. Soln of 5-7% Widely used as secondary fixative Black depositsremove by adding sat. iodine soln in 96% alcohol Mercuric chloride stock soln + glac. HoAcprevent turbidity and formation of dark ppt

Glutaraldehyde

hour (small specimen) to 2 hours (maximum contribution)

Metallic Fixatives: A. Mercuric chloride

Mercuric chloride

Mercuric chloride [5gm] Potassium dichromate [2.5gm] Sodium sulfate (optional) [1gm] Distilled water [100gm] Mercuric chloride stock [95ml] Glac. HoAc [5ml] (to be added just 12-24 hours

Zenkers fluid

before use

12-24 hours

Zenker-formol (hellys solution)

Mercuric chloride, stock solution: Mercuric chloride [5gm] Potassium dichromate [2.5gm] Sodium sulfate [1gm] Distilled water [100ml] Strong formaldehyde, 40% [5ml]to be added just before use

Compatible woth most stains Not stable until after addition of HoAc Causes lysis of RBCs and removes iron from hemosiderin De-zenkerizationremoval of mercuric deposits by alcoholic iodine soln Excellent microanatomic fixative for pituitary gland, bone marrow, and blood containing organs such as spleen and liver.

B. Chromate fixative

C. Lead fixative