Error Status code: 401
OPenT (Optical Projection Tomography).
Location :
Microscope: Custom made body
Year Installed: 2014
Camera: Hamamatsu Flash Orca 4.0 LT 2048*2048*
Optics: Fluorescence and Transmitted light
Operating: BABB or other clearing protocols2)
* - camera may be working at a lower format.
Keywords:
Text still being written.
Suggestion for description in "Materials and Methods":
Sample material and methods text under construction.
We would like to thank the technical support of IGC's Advanced Imaging Facility (AIF-UIC), which is supported by the national Portuguese funding ref# PPBI-POCI-01-0145-FEDER-022122, co-financed by Lisboa Regional Operational Programme (Lisboa 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER) and Fundação para a Ciência e a Tecnologia (FCT; Portugal).
Configuration |
Value |
Below is a list of the objectives and filters available. On the DeltaVision system, the filter cube (dichroic) is manually set and the emission filters are split between two filter wheels that also need to be manually switched. Make sure that both match the excitation and emission filters you want to use.
Magnification |
Objective type |
NA1 |
Immersion |
Working distance (mm) |
Resolution2 @525nm in XY |
Image Pixel size3 |
Image Pixel size SIM3 |
Link |
|
20x ![]() |
UPlan SAPO |
0.75 |
- |
0.60 |
0.427 |
0.240 |
0.120 |
||
60x ![]() |
Plan Apo N |
1.42 |
OIL |
0.15 |
0.226 |
0.080 |
0.040 |
(1) NA - Numerical Aperture. For more information, follow this link.
(2) These values are the theoretical optical resolution limits depending on the NA of the objective and were calculated using the Resolution formula adapted to fluorescence as seen in the Olympus micro website: R = 0.61*λ/NA
(3) The pixel size saved automatically with the .dv files are estimates. The values on this table were measured with a stage graticule, with and without the 1.6x auxiliary magnification. For more accurate measurements use these values instead.
IMPORTANT: For optimal deconvolution you may use the 1.6x auxiliary magnification, however, using this may cause aberration on the edges of the field of view!
Fluorescence filterset |
Emission wavelength (λ) |
Description |
Camera |
DAPI |
435/31 |
DAPI, Hoesche, etc. |
1 |
GFP |
528/48 |
GFP, FITC, Alexa Fluore 488, etc. |
1 |
RFP |
609/37 |
RFP, TRITC, mCherry, AF568, etc. |
2 |
Cy5 |
683/40 |
Far Red, Cy5, AF633, AF647, etc. |
2 |
DIC |
- |
Transmitted Light |
1/2 |
The DeltaVision tends to create large files because of the capability to perform live imaging with multiple channels and multiple positions. As such it has been setup as an FTP server so users can download the files from the comfort of their desk. Learn how to below.
Quick connection, fill the top bar:
Host:
username & password
If you want to learn more timesaving features of FileZilla, visit their website.
ImageJ/Fiji recognize and open DeltaVision .dv files, but you will need to use the Bio-Formats plugin. This plugin is available by default in the Fiji distribution.
To install the plugin in ImageJ, download the latest stable release of Bio-formats and drag it into the plugins folder. Once you start ImageJ again it will appear on the plugins menu. Go to "Plugins>Bio-formats>Bio-formats importer" or drag-and-drop a file in ImageJ/Fiji to open it.
Differential Interference Contrast (DIC)