NIH 3T3 cell expressing EGFP-Lifeact (a small genetically expressed probe that binds to actin and is derived from the first 17 aa of Abp140) imaged with structured illumination. Structured illumination produces images that have a lateral resolution of approximately 100 nm, or twice the diffraction limited 200 nm resolution of conventional fluorescent microscopy techniques. There are three related images in this folder. This file is the raw data. The 5 positions of the illumination grid pattern are visible in the 10 slices of the z-series. The structure of this file is that all 150 images appear as a horizontally incrementing sequence. The second file is a z-series of six of the reconstructed structured illumination images. The third file contains the maximum projection image of the second file. The second and third files each have 4 channels of data to compare image resolution: channel 1 - structured illumination, channel 2 - wide field, channel 3 - deconvolved wide field, and channel 4 - filtered deconvolved wide field. Images were collected with a Carl Zeiss SIM microscope using a 63 X 1.4 Plan Apo objective, an Andor Ixon camera with 150 ms exposures, Zeiss filter set 2, and a 488 nm laser.
Spatial Axis | Image Size | Pixel Size |
---|---|---|
X | 1004px | 0.04µm |
Y | 1002px | 0.04µm |
Z | —— | 0.12µm |