Human Renal Nerve Study Plan 9.10.08

1. Evaluate entire section. Exclude any samples that are incomplete. 2. Count nerves per the following size criteria (measure smallest diameter):

a. Small (S) is < 0.2 mm b. Medium (M)is 0.2 to <0.5 mm c. Large (L) is 0.5 to <1.0 mm d. Extra Large (XL) is ≥ 1.0 mm

3. For each nerve count and capture distance from IEL (if no IEL is visible, estimate the location of IEL and measure from that location)

4. If there is significant disease, please comment on disease 5. After counting the nerves in a given section, comment on the distribution. If there is a given

region with a greater concentration of nerves, comment on which general area the greatest concentration of nerves fall

6. Capture one perfusion fixed renal artery block 7. Would like to send one set of samples for S-100 staining. Either it can be done at Stanford, or if

this is not easy, we can send it to a lab that we work with

• Working Strategies:• • Define all zones on slides, mark with dot lines• Take a low magnification (1.5x) of each section for dimension measurement in the future• Take image(s) at 5x for each zone, may require more than 1 images, stitch them• Verify each nerve under higher magnification, and• Mark each nerve with number on images, and count them for each zone• Measure diameter, distance from IEL on the images• Record data on xls

Define all zones on slides, mark with dot linesTake a low magnification (1.5x) of each section for dimension measurement in the future

Take image(s) at 5x for each zone, may require more than 1 images, stitch them. Nine for the superior zone of sextion 33276_LRA_Prox_1A

Verify each nerve under higher magnification, andMark each nerve with number on images, and count them for each zone

Click on the image to see the larger image for each nerve. The file is pretty large and may be saved on pc for better browsing.

Measure diameter of the nerves on the images

Click on the image to see the larger image for each nerve. The file is pretty large and may be saved on pc for better browsing.

Measure distance of the nerves from IEL on the images

Click on the image to see the larger image for each nerve. The file is pretty large and may be saved on pc for better browsing.

Record data on xls Click on the image to open the data.xls fileThis file is still a draft…

• Please advise

• Thanks!