Oliver Smithies:
We go to book T starting on Tuesday, April 26th and running to November. I remember this period of my career as being neither very enjoyable nor productive. Starting the beginning with neonatal lymph node. C57 black delivers a large litter. And india ink to get the lymph node. Rather unsuccessful. Fixation test. Lymph node staining. [00:01:00] India ink plus or minus saline on lymph node. This sort of experiment going on. Vascular picture plus or minus antigen on page 11 May 2nd. Trying to get a vascular pattern of some sort. [00:02:00] More pages of presumably Alice’s or maybe [Nancy’s?] writing April 29th, Friday, page 15. A gel. Might have been mine, might not have been. Probably not. Two urine samples and one serum sample from Dr. Clatanoff. This is Tuesday, May 10th. Not in my handwriting. And with results shown on the left. [00:03:00] Urine and the plasma. Back to india ink and antigen tests in my writing Wednesday, May 18th. Fixation difficulties. Conclusions of fixatives on page 27. [00:04:00] Tests of — I think these are brucella titrations from the look of it on page 35, May 27th. Yes, Brucella abortus antigen. And following pages many images in 96-well plates. What the pellets looked like. Eventually that type of experiment was made very sensitive by myself and Tom Wegmann. This we will maybe find out later. Wednesday, June 1st, repeat of the Brucella abortus series and extensions to looks like Salmonella Pullorum. [00:05:00] Stained antigens available I suppose.
Ring test antigens Tuesday, Wednesday, June 1st, continuing on the next few days. Some TVB gels of multiple myeloma June 22nd. My notes again, Wednesday, August 17th. Fluorescein diacetate for [00:06:00] cell labeling, etc. Trying to make acetate. That’s on Thursday, August 19th, gram of fluorescein with 5 ml of acetic anhydride to make fluorescein acetate as a substrate for tests. Organic chemistry continuing on page 57 and on page 56. [00:07:00] Melting point 203 degrees centigrade with a Japanese thermometer. The crystalline ferrocene acetate. Tested 1 microliter with 1,000 microliters of medium (inaudible) works beautifully. I don’t know what work is. But that said it worked beautifully. We began to use an Autotechnicon system for fixing things. [00:08:00] Began to see this Tuesday, August 26th, page 61. Into the Autotechnicon fixative about 3:00 p.m., etc.
Autotechnicon-fixed and stained with a note on page 60. The mitoses are good. Sunday, September 4th (inaudible) final stain, a modified Martius yellow solution, etc. This is it. I must have liked the stain. [00:09:00] Frozen section. Frozen slide fixes, etc. Trying the staining again looking at mitoses and deciding whether they were good to see.
Still not at all clear what the motivation of these experiments was. [00:10:00] India ink again on Wednesday, September 14th. C57 black with Nembutal. India ink into the heart after pulling out a little, etc. Fluorescein diacetate continued September 15th. [00:11:00] Working quite a long time on this problem. Page 95, September 17th, Saturday (inaudible) try isopentane, etc. The frozen dried sections in paraffin. Spreading not possible on water, etc., etc. Typical experiment here of testing [00:12:00] an enormous number of solvents. Chloroform, carbon tetrachloride, butyl Cellosolve, ethyl ether, cyclohexane, hexanone, acetone, toluene, dimethyl sulfoxide, [formicin?], diacetone alcohol, benzyl alcohol, etc., etc. Butyl alcohol at 47 degrees promising. All on one page. Long term dry DMSO lost its fluorescence. Butyl alcohol and diacetone alcohol did not. But did not dry, etc., etc. [00:13:00] Real fluorescein diacetate test on Saturday, September 24th, page 103. Ed Azen and I sent a letter to Dr. [Russ Langley?] on page 105 summarizing what we’d done or what he’d done with erythrocyte stroma and gel electrophoresis. Which we [00:14:00] did publish a paper, Ed and I, on this topic, which I’ll find in a moment. We published a paper on starch gel electrophoresis of erythrocyte stroma which was submitted August 18th, 1964. So that’s — I’ve lost a year.
Interviewer:
Sixty-six.
OS:
This book. What book is this?
I:
Sixty-six.
OS:
So this is quite a long time after the publication that Ed and I had in 1964. It’s just a letter to help somebody else who wanted to know. But it’s rather big. It’s a rather nice summary of what Ed Azen did. [00:15:00] Are we still —
I:
It is.
OS:
It’s still running. Yeah, OK, good. Fluorescein acetate labeling again. October 10th. Evidently motivated in some way by Gowans’s experiments because this is on October 12th, page 115, Wednesday. There’s continue and use Gowans’s conditions. [00:16:00] I know I’d been teaching Gowans’s work in recirculation of the lymphocytes.
Had some problems with staining and getting crystals. Evidently caused some problem because there’s a whole page on 119 on crystal removal with treating in certain way. No crystal with three exclamation marks. Messing around with stains. Here on the following page is beginning to show [00:17:00] the thymus shielding again. How the lead could be arranged before irradiation. Trying to get autoradiography to work with the help of [Walter Plow?] evidently on page 127. [00:18:00] Cultures continued Friday, November 11th. Trying to culture them on top of a filter that allowed nutrient to come from the bottom but the cultures were on top of a so-called [grove stein?] dish. [00:19:00] Some photographic images on page 138 and 139. The control in 1 hour and 18 hours and so on. Not much guide as to what these mean.
The following pages. Cultures continue Thursday, November 17th. Decided to [set?] the concentration of serum and the size of the explant. Trying to culture the thymus cell. [00:20:00] With the results. The results suggest that large pieces are as good as or better than small. But the histology is poor.
On the following pages are many images of the cultures. Viable cortex and dead cortex and so on. Continuing to struggle with it. Cultures again Wednesday, November 23rd. With a [sieve three?] micron membrane, etc. [00:21:00] Totally submerged cultures versus on top. Try adding red cells, so forth. And images again of the thymus for the following pages that end this book. T.