Confocal Microscopy Analysis of Intestinal Tissues

Tissues were analyzed by cryostat section staining using confocal microscopy as previously described [5 (link)]. Jejunum and colon tissues were double stained with anti-SRF (1:500, Santa Cruz Biotechnology, Dallas, TX) and anti-αSMA (1:1200, Sigma, St. Louis, MO) antibodies. Stained tissues were analyzed using confocal microscopy. For histological analysis, tissues were dehydrated, embedded in paraffin, cut into 4 μm-thick coronal sections, rehydrated and stained with H&E. Images were collected using an Olympus FV1000 confocal laser scanning microscope with Fluoview FV10-ASW 3.1 Viewer software (Olympus, Tokyo, Japan) or the iScan Coreo scanner (Ventana Medical Systems, Tucson, AZ).

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Lee M.Y., Park C., Ha S.E., Park P.J., Berent R.M., Jorgensen B.G., Corrigan R.D., Grainger N., Blair P.J., Slivano O.J., Miano J.M., Ward S.M., Smith T.K., Sanders K.M, & Ro S. (2017). Serum response factor regulates smooth muscle contractility via myotonic dystrophy protein kinases and L-type calcium channels. PLoS ONE, 12(2), e0171262.

Publication 2017

anti-SRF anti-αSMA FV1000 confocal laser scanning microscope Fluoview FV10-ASW 3.1 Viewer software iScan Coreo scanner

Antibodies Colon Confocal laser scanning microscope Embedded paraffin Jejunum Stained tissues Tissues αsma

Corresponding Organization : University of Rochester

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Variable analysis

independent variables

Tissue type (jejunum and colon)

dependent variables

Protein expression of SRF and αSMA
Histological analysis of tissue samples

control variables

Staining and microscopy protocols as previously described [5]
Antibody concentrations (anti-SRF 1:500, anti-αSMA 1:1200)

controls

Positive control: Double staining with anti-SRF and anti-αSMA antibodies
Negative control: Not explicitly mentioned

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