Life Sciences Commons^™

Gpu Deconvolution Wow Result On 2048x2048x32 Plane Z-Series, George Mcnamara

George McNamara

GPU Deconvolution WOW result on 2048x2048x32 plane Z-series ... formerly bad academic code ("you get what you pay for") now impressive

Alternative title: "instant gratification quantitative deconvolution fluorescence microscopy".

http://works.bepress.com/gmcnamara/55/

Please see "74"

http://works.bepress.com/gmcnamara/74/

for 32-bit images from this project (bepress file size limitation prevented me from including them in this ZIP archive).

//

Summary: Deconvolution microscopy has historically been painfully slow. The early vendors were:

- Scanalytics (Carrington and Fay), commercialized to try to sell expensive, specialized array processors made by CSPI (the CSPI box likely had less computing power than a first gen smartphone).

- Applied Precision (Sedat …

More Fish (1 Of 4), George Mcnamara

George McNamara

More Stellaris FRISH and GPU Deconvolution, 20140313Thur - part 1 of 4

Please see other content in the FISH Imaging section of

http://works.bepress.com/gmcnamara/subject_areas.html

for details on the microscope. Briefly: Leica DMI6000 inverted microscope, SOLA lamp, Hamamatsu FLASH4.0 sCMOS camera, 100 nm XY, 200 nm Z pixels, 32 planes (16 um Z), MetaMorph 7.8 acquisition software.

Four zip files:

http://works.bepress.com/gmcnamara/43

http://works.bepress.com/gmcnamara/44

http://works.bepress.com/gmcnamara/45

http://works.bepress.com/gmcnamara/46

with two dataset (001, 002), raw and deconvolved using Bruce & Butte GPU deconvolution in Fiji ImageJ,

http://www.ncbi.nlm.nih.gov/pubmed/23482010

http://www.opticsinfobase.org/vjbo/fulltext.cfm?uri=oe-21-4-4766&id=249375 (free download, as are all Optics Express papers).

http://tcell.stanford.edu/software.html

http://lists.umn.edu/cgi-bin/wa?A2=ind1312&L=confocalmicroscopy&P=5534

Note: "001 raw" (bepress 43) contains my "imaging SOP" …

More Fish (2 Of 4), George Mcnamara

George McNamara

More Stellaris FISH and GPU Deconvolution, 20140313Thur - part 2 of 4

Please see other content in the FISH Imaging section of

http://works.bepress.com/gmcnamara/subject_areas.html

for details on the microscope. Briefly: Leica DMI6000 inverted microscope, SOLA lamp, Hamamatsu FLASH4.0 sCMOS camera, 100 nm XY, 200 nm Z pixels, 32 planes (16 um Z), MetaMorph 7.8 acquisition software.

Four zip files:

http://works.bepress.com/gmcnamara/43

http://works.bepress.com/gmcnamara/44

http://works.bepress.com/gmcnamara/45

http://works.bepress.com/gmcnamara/46

with two dataset (001, 002), raw and deconvolved using Bruce & Butte GPU deconvolution in Fiji ImageJ,

http://www.ncbi.nlm.nih.gov/pubmed/23482010

http://www.opticsinfobase.org/vjbo/fulltext.cfm?uri=oe-21-4-4766&id=249375 (free download, as are all Optics Express papers).

http://tcell.stanford.edu/software.html

http://lists.umn.edu/cgi-bin/wa?A2=ind1312&L=confocalmicroscopy&P=5534

Note: "001 raw" (bepress 43) contains my "imaging SOP" …

More Fish (3 Of 4), George Mcnamara

George McNamara

More Stellaris FISH and GPU Deconvolution, 20140313Thur - part 3 of 4

Please see other content in the FISH Imaging section of

http://works.bepress.com/gmcnamara/subject_areas.html

for details on the microscope. Briefly: Leica DMI6000 inverted microscope, SOLA lamp, Hamamatsu FLASH4.0 sCMOS camera, 100 nm XY, 200 nm Z pixels, 32 planes (16 um Z), MetaMorph 7.8 acquisition software.

Four zip files:

http://works.bepress.com/gmcnamara/43

http://works.bepress.com/gmcnamara/44

http://works.bepress.com/gmcnamara/45

http://works.bepress.com/gmcnamara/46

with two dataset (001, 002), raw and deconvolved using Bruce & Butte GPU deconvolution in Fiji ImageJ,

http://www.ncbi.nlm.nih.gov/pubmed/23482010

http://www.opticsinfobase.org/vjbo/fulltext.cfm?uri=oe-21-4-4766&id=249375 (free download, as are all Optics Express papers).

http://tcell.stanford.edu/software.html

http://lists.umn.edu/cgi-bin/wa?A2=ind1312&L=confocalmicroscopy&P=5534

Note: "001 raw" (bepress 43) contains my "imaging SOP" …

More Fish (4 Of 4), George Mcnamara

George McNamara

More Stellaris FISH and GPU Deconvolution, 20140313Thur - part 4 of 4

Please see other content in the FISH Imaging section of

http://works.bepress.com/gmcnamara/subject_areas.html

for details on the microscope. Briefly: Leica DMI6000 inverted microscope, SOLA lamp, Hamamatsu FLASH4.0 sCMOS camera, 100 nm XY, 200 nm Z pixels, 32 planes (16 um Z), MetaMorph 7.8 acquisition software.

Four zip files:

http://works.bepress.com/gmcnamara/43

http://works.bepress.com/gmcnamara/44

http://works.bepress.com/gmcnamara/45

http://works.bepress.com/gmcnamara/46

with two dataset (001, 002), raw and deconvolved using Bruce & Butte GPU deconvolution in Fiji ImageJ,

http://www.ncbi.nlm.nih.gov/pubmed/23482010

http://www.opticsinfobase.org/vjbo/fulltext.cfm?uri=oe-21-4-4766&id=249375 (free download, as are all Optics Express papers).

http://tcell.stanford.edu/software.html

http://lists.umn.edu/cgi-bin/wa?A2=ind1312&L=confocalmicroscopy&P=5534

Note: "001 raw" (bepress 43) contains my "imaging SOP" …

Time Series Data For 3b Image Processing, George Mcnamara

George McNamara

Time series data for 3B image processing

Four time series data sets of Stellaris single molecules RNA FISH (fluorescence in situ hybridization). All four datasets are 301 imaqe planes. FISH probes are to TOP1 mRNA (Topoisomerase I)in Saos osterosarcoma cells.

One dataset is 10 millisecond exposures (very dim), acquired in Streaming acquisition mode (no hardware overhead).

The other three datasets are a three consecutive planes of the same XY field of view.

I have included image processing results for:

PiMP - a fast method developed by Sebastian Munck et al 2012, http://www.ncbi.nlm.nih.gov/pubmed/?term=22357945

and greatly improved performance by Glen Macdonald (Seattle). …

Stellaris Fish Workshop Gadph And Dapi Z-Series, George Mcnamara

George McNamara

Stellaris FISH workshop GADPH and DAPI Z-series at MD Anderson Cancer Center, Houston, TX.

The zip file contains raw and GPU deconvolved image data from a workshop Biosearch Technologies conducted for MDACC researchers in December 2013. Image data was acquired on a Leica DMI6000 microscope with Lumencor SOLA light engine, DAPI and Cy5 filter cubes, Hamamatsu ORCA FLASH4.0 sCMOS camera (500 ms exposure time per plane for Quasar 670).

Pixel size 100 nm XY.

Z-step size 200 nm.

32 planes (power of 2 is optimal for GPU deconvolution). With 500 ms exposure time, the Quasar 670 GADPH FISH probes images …

Stellaris Fishing 20131125mon Part 1 Of 2, George Mcnamara

George McNamara

Stellaris FISH dataset using three FISH probe sets. Slides courtesy of Biosearch Technologies,

https://www.biosearchtech.com/store/product.aspx?catid=224,318,324

see http://stellarisfish.smugmug.com/ for online gallery by Biosearch.

This experiment was to evaluate the crosstalk between the Biosearch fluorophores:

Quasar 570

CAL Fluor Red 610 (CFR 610)

Quasar 670

DAPI (DNA counterstain)

Autofluorescence (green, but sometimes showing up in other channels).

and our lab's Leica DMI6000 fluorescence microscope with Leica filter sets:

DAPI

GFP (L5)

Cy3 (N3)

Texas Red (TxRed2)

Cy5 (Y5)

I also acquired green channel and red channel with exciter filters in our ASI excitation wheel:

GFP + 492 exciter

Texas Red (TxRed2) + 572 …

Stellaris Fishing 20131125mon Part 2 Of 2, George Mcnamara

George McNamara

Stellaris FISH dataset using three FISH probe sets.

Slides courtesy of Biosearch Technologies,

https://www.biosearchtech.com/store/product.aspx?catid=224,318,324

see http://stellarisfish.smugmug.com/ for online gallery by Biosearch.

This experiment was to evaluate the crosstalk between the Biosearch fluorophores:

Quasar 570

CAL Fluor Red 610 (CFR 610)

Quasar 670

DAPI (DNA counterstain)

Autofluorescence (green, but sometimes showing up in other channels).

and our lab's Leica DMI6000 fluorescence microscope with Leica filter sets:

DAPI

GFP (L5)

Cy3 (N3)

Texas Red (TxRed2)

Cy5 (Y5)

I also acquired green channel and red channel with exciter filters in our ASI excitation wheel:

GFP + 492 exciter

Texas Red (TxRed2) + 572 …

Leica Microscope Gpu Deconvolution Stellaris Fish Dataset #1, George Mcnamara

George McNamara

McNamara 20131101Fri Leica widefield microscope CUDA Deconvolution Stellaris FISH probe cultured cells dataset #1.zip

A text file in the zip archive has experiment details. I am posting this online so that researchers - whether academic or commercial - can evaluate the acquired data, the Bruce and Butte 2013 Optics Express ( http://www.opticsinfobase.org/oe/fulltext.cfm?uri=oe-21-4-4766 ) deconvolution result (note: I may not have used optimal settings), and to compare these deconvolution results to other methods. If anyone generates alternative spatial deconvolution output, such as from: * SVI Huygens * Media Cy AutoQuant * Agard's ER-Decon (Arigovindan 2013 PNAS) * Vicidomini SGP GPU Deconv …