Re: [Histonet] insect histology book inquiry

From:Philip Oshel


"Stiles' n-butyl alcohol technique
Recommended as particularly useful for small insects.
Solution A
  ethyl alcohol (45%)  90 pt
  n-butyl alcohol         10 pt

Solution B
  ethyl alcohol (62%)  80 pt
  n-butyl alcohol         20 pt

Solution C
  ethyl alcohol (77%)  65 pt
  n-butyl alcohol         35 pt

Solution D
  ethyl alcohol (90%)  45pt
  n-butyl alcohol         55 pt

Solution E
  ethyl alcohol (abs)  2 pt
  n-butyl alcohol        1 pt

Solution F
  Paraffin (m.p. 56-58 deg C)   2 pt
  n-butyl alcohol        1 pt

1. After fixation (e.g. Gilson's) place in ethyl alcohol (35%) for 30 
min. to 1 hr.
2. Transfer to solution A for 2 hr.
3. Transfer to solution B for 2 hr.
4. Transfer to solution C for 4 hr.
5. Transfer to solution D for 6 hr. to several days.
6. Transfer to solution E for 6 hr. to overnight (with one change)
7. Transfer through 2 changes of n-butyl alcohol (at several hours interval).
8. Transfer to solution F in covered dish (in oven) for 12 to 24 hr.
9. Uncover, dish until n-butyl odor dissipates.
10. Transfer to paraffin (optional).
11. A long infiltration period (4-5 days) is recommended for best 
results (causes less hardening).
12. Embed in paraffin wax (56-58 deg C m.p.) with slight addition of 
bayberry wax (<=3%).

1. The choice of fixative is important so that no hardening factors 
are added to the procedure.
2. Specimens may be left in n-butyl (step 7) for a very long period of time."

ref. cited:
Stiles, K.A. 1934. Normal butyl alcohol technic for animal tissues 
with special reference to insects. Stain Technol. 9:97-100.

Barbosa, P. 1974. Manual of Basic Techniques in Insect Histology. 
Autumn Publishers, Amherst, MA.

Sounds like a relaxing technique.


>Good Thursday morning!
>Is there anybody out there in histoland who might have a copy of the 
>now out of print book, A Manual of Basic Techniques in Insect 
>Histology by P. Barbosa, Autumn Publishers, 1974?
>I'm looking for details on Stile's butyl alcohol technique that was 
>discussed in a paper on whiteflies and Barbosa was cited.
>A jillion thanks to anybody who can provide this technique!
>Andi Grantham
>: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
>: Sr. Research Specialist       University of Arizona               :
>: (office:  AHSC 4212)          P.O. Box 245044                     :
>: (voice:  520-626-4415)        Tucson, AZ  85724-5044    USA       :
>: (FAX:  520-626-2097)          (email:       :
>Histonet mailing list

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
024C Brooks Hall
Mt. Pleasant, MI 48859
(989) 774-3576

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