Dataset from "Channelrhodopsin-2 Oligomerization in Cell Membrane Revealed by Photo-Activated Localization Microscopy" (b1AR-mEos3.2)

<p>This dataset contains raw microscopy data from the article "Channelrhodopsin-2 Oligomerization in Cell Membrane Revealed by Photo-Activated Localization Microscopy" (doi.org/10.1002/anie.202307555)</p> <p>Zip file contains PALM measurements of HEK293 cells expressing &szlig;₁AR fused with mEos3.2.<br>Each folder in .zip file contains one PALM measurement (TIFF format) and recording settings (.xml) automatically created by acquiring software. For technical reasons, single measurements are divided into TIFF files of =&lt;4088 frames.</p> <p>Detailed protocols for sample preparation and data acquisition are described in the article.<br>Some details are listed below.</p> <p><strong>SAMPLE PREPARATION</strong></p> <p>cell line: Flp-In&trade; T-REx&trade; 293<br>transfection method: modified calcium-phosphate transient transfection [1]<br>fixation method: 4% paraformaldehyde solution in PBS for 30 min at room temperature<br>imaging buffer: PBS</p> <p><strong>PALM ACQUISITION</strong></p> <p>A custom-built setup for single-molecule localization microscopy is described in [2].<br>mEos3.2 was simultaneously photoconverted, imaged and photobleached by gradually increasing UV illumination (405 nm; up to 1 mW at the sample) and continuous excitation at 561 nm (approximately 50 mW at the sample).<br>PALM movies of cell plasma membrane were recorded in total internal reflection fluorescence (TIRF) mode using an EMCCD camera:<br>exposure time = 100 ms<br>frame size = 512x512 pixels<br>pixel size = 80 nm</p> <p>&nbsp;</p> <ol> <li>Chen C, Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Molecular and cellular biology. 1987 Aug 1;7(8):2745-52.</li> <li>Tang Y, Dai L, Zhang X, Li J, Hendriks J, Fan X, Gruteser N, Meisenberg A, Baumann A, Katranidis A, Gensch T. SNSMIL, a real-time single molecule identification and localization algorithm for super-resolution fluorescence microscopy. Scientific reports. 2015 Jun 22;5(1):11073</li> </ol>