1. IF-LCM: Laser capture microdissection of immunofluorescently defined cells for mRNA analysis 
Hiroshi Murakami, Lance Liotta, Robert A. Star 
Kidney International 
Volume 58, Issue 3, Pages (September 2000)
DOI: /j x
Copyright © 2000 International Society of Nephrology Terms and Conditions

2. Figure 1
Time course of RNA disappearance by acridine orange. Sections of mouse kidney were fixed in ice-cold acetone, incubated in diethel pyrocarbonate (DEPC)-treated water for 1, 3, 5, or 10 minutes at room temperature, and then rapidly stained with 0.02% acridine orange. After a brief washing, the sections were examined using an ultraviolet filter cube, and the image was captured with a Coho video camera.
Kidney International  , DOI: ( /j x)
Copyright © 2000 International Society of Nephrology Terms and Conditions

3. Figure 2
Time course of RNA disappearance of reverse transcription-polymerase chain reaction (RT-PCR). Cryosections (2 μm) of a mouse kidney were exposed to 0 (control), 1, 3, 5, or 10 minutes of DEPC-treated water at room temperature, dehydrated, and air dried, and then manually scraped with a clean razor blade as described previously6. The RNA was extracted, and RT-PCR was performed for malate dehydrogenate (MDH). Columns are successive 1:10 dilutions of cDNA obtained from the entire section. MDH mRNA was not detected in the one-minute sample processed without RT (data not shown).
Kidney International  , DOI: ( /j x)
Copyright © 2000 International Society of Nephrology Terms and Conditions

4. Figure 3
Laser capture microdissection capture of immunofluorescently stained mouse kidney. A 2 μm section of freshly frozen mouse kidney tissue was fixed in 100% acetone and rapidly immunostained with anti-Tamm-Horsfall protein (anti-THP; Methods section). (A and B) The immunofluorescent and transmitted light image before transfer, respectively. (C) The fluorescent image of the tissue section after dissection. (D) A fluorescent image of the thick ascending limb (TAL) recovered on the transfer film. The arrow indicates the transferred area.
Kidney International  , DOI: ( /j x)
Copyright © 2000 International Society of Nephrology Terms and Conditions

5. Figure 4
Specificity of IF-LCM capture. RT-PCR analysis of three genes in TAL sample isolated by LCM. Mouse outer medullary TALs were labeled with THP as in Figure 3. All the TALs on a single kidney cross-section were microdissected using approximately μm diameter laser spots. Each RT-PCR reaction used 7% of the RNA obtained from all of the TALs identified on a single kidney cross-section. Table 1shows the details of PCR primers and reaction conditions. Abbreviations are: rBAT, basic amino acid transporter; THP, Tamm-Horsfall protein; AQP-2, aquaporin-2. Positive control (total), pooled sample of RNA harvested from scraped section from same block. Negative control [RT(-)], total RNA from microdissected TAL analyzed without reverse transcriptase.
Kidney International  , DOI: ( /j x)
Copyright © 2000 International Society of Nephrology Terms and Conditions

6. Figure 5
mRNA recovery following immunofluorescent (IF) and hematoxylin and eosin (H&E) staining. Serial 2 μm cryosections of mouse kidney were fixed with acetone and dehydrated (without exposure to aqueous solutions, control), rapidly immunofluorescently labeled with THP, or stained with HE. Slides were scraped. RNA was extracted, and RT-PCR was performed for MDH mRNA as in Figure 2. Each RT-PCR reaction tube used 7% of the RNA obtained from a single 2 μm cryosection.
Kidney International  , DOI: ( /j x)
Copyright © 2000 International Society of Nephrology Terms and Conditions