LESSON 16: ADHESIVES AND MOUNTING MEDIA

 After cutting, sections are floated out on a water bath.

 When the sections chosen have flattened out, the numbered slide is immersed in the water bath and the section
is fished out and drained.
 After draining the sections are fixed to the slide.
This can be done either by : leaving the slides in a 37C incubator over night, by placing the slides in a wax oven
at 56 – 60C for 2 hours, or by drying the slides on a hot plate at 45 – 55C for 30 to 40 minutes.
 Another alternative to drying is by the use of adhesives.
 To promote adhesion of sections, adhesive may be spread thinly and evenly on a clean grease free slide which
is then gently approximated to the end of the ribbon, and drawn upwards in a near vertical motion.

ADHESIVE
 Not necessary for routine staining, provided than the slides are clean and free from grease
 However they are essential for methods that require exposure of sections to acids and alkalis during staining.
 Alternative to drying
 Smeared to the slides so that the sections stick well to the slides

The Common Adhesive used are the following:

 Mayer’s albumin
 MOST commonly used because very easy to make and is convenient and inexpensive.
 A drop of Mayer’s Egg Albulin is usually smeared into the clean glass slide before sections are oriented.
 Dried albumin
 Crystals of thymol is added
 Componnents: Dried albumin, NaCl
 Gelatin (1%)
 Added and mixed to the water bath
 Most convenient alternative to direct coating of slides
 Gelatin-formaldehyde mixture
 Coated slides are dried at 37C for 1 hour or overnight before use
 Components: 1% gelatin and 2%formaldehyde
 Starch paste
 Thymol crystals are added to prevent mold formation
 Components: Powdered starch, dH20 and HCl
 Plasma
 Readily available from outdated blood
 Dispensed into sterile tubes (0.5mL)
 Poly-L-Lysine
 Widely used section adhesive in immunohistochemistry
 APES (3-aminopropylthriethoxysilane)
 Are very useful in cytology, particularly in cytospin preparations of proteinaceous or bloody material.

MOUNTING MEDIUM
 Usually a syrup fluid applied between section and the coverslip after staining, setting the section firmly,
preventing the movement of the coverslip
 It protects the stained section from getting scratched, and from bleaching or deterioration due to oxidation,
thereby preserving the slides for permanent keeping, to facilitate easy handling and storage.
 It also serves to prevents the distortion of image during microscopic examination.
PROCEDURE:
 The slide carrying the section to be mounted is taken from the last xylene bath with the forceps.
 A drop of mounting medium is placed down the center of the slide.
 A celan, dry coverslip is placed on the slide.
 The slide may then be incubated at 37C for 12-24 hours after mounting, to harden the medium.
 Characteristics of a Good Mounting Medium
 The refractive index of the mountant should be near to that glass which is 1.518
 Freely miscible with xylene and toluene
 Should not dry quickly
 Should not crack or produce artefactual granularity on the slide upon drying
 Should not dissolve out or fade tissue sections
 Should not cause shrinkage and distortion of tissues
 Should not bleach out any stain or affect staining
 Should not change in color or pH
 Should not set hard, thereby producing permanent mounting of sections

Mounting media may be divided into two main groups:

 Aqueous media
 Resinous media

Aqueous Mounting Media

 Designed to mount water miscible preparation directly from water in cases where the stain removed or
decolorized with alcohol or xylene
 Made up of gelatin, glycerine jelly or gum arabic (to solidify the medium), glycerol(prevent cracking and
drying of the preparation) , sugar ( increase the refractive index)and preservative solution.

EXAMPLES of Aqueous Mounting Media

A. Water
 Has low refractive index, moderately transparent and evaporates easily, hence is good only for temporary
mounting.
 Does not allow tissues to be examined under the oil immersion lens.

B. Glycerin
 May also be used as preservative, has high refraction index and lasts for few minutes
 Provides greater visibility if slightly diluted with water
 Suitable semi-permanent mounting medium with a refractive index of 1.46, has preservative action, sets quite
hard, and will keep sections mounted for years, especially if sealed on the edges with paraffin.

C. Farrant’s medium
 RI.: 1.43
 It takes longer time to harden .

D. Apathy’s medium- 1.52

Used for methylene blue stained nerve preparations.

E. Brun’s fluid- for frozen sections

F. Karo Corn syrup

Resinous Mounting Media
 Used for preparation that have been dehydrated and cleared in xylene or toluene, and are recommended for
majority of staining methods
 They are subdivided into natural and synthetic resins

EXAMPLES Resinous Mounting Media

A. Canada balsam
 Natural resin extracted from the Canadian tree, Abus Balsamea.
 Dissolved in xylene in an incubator at 37C or paraffin oven at 58C, and filtered, obtaining the desired
consistency by controlled evaporation of the solvent.
 Transparent almost colorless oleoresin that adheres firmly to glass and sets to a hard consistency without
granulation
 Recommended for whole mounts and for thick sections because it does not shrink much
REFRACTIVE INDEX: 1.524

B. Eukitt

C. Etallan

D. DPX
 This is a resinous medium recommended for small tissue sections but not for whole mounts because of
shrinkage produce on drying.
REFRACTIVE INDEX: 1.532

E. Histomount

F. XAM
 It is a synthetic resin mixture in xylene.
 Dries quickly and preserves stains well.
REFRACTIVE INDEX: 1.52

H. Clarite
AKA clarite X.
It is a synthetic resin which is soluble in xylene
It is generally preffered over D.P.X.
REFRACTIVE INDEX: 1.544

Other recommended synthetic media:

 Permount
 Harleco Synthetic Resin
 Clearmount

RINGING
 It is the process of sealing the margins of the coverslip to prevent the escape of fluid or semi fluid mounts and
evaporation of mountant, to immbolized the coverslip, to prevent sticking of the slides upon storage.
Examples of Ringing Media:
 Kronig Cement
 Durofix