imagestreamx
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| - | <img src="/facility/flowcytometry/lib/exe/fetch.php?media=imagestreamx_name.jpg"> | + | <img src="/facility/flowcytometry/lib/exe/fetch.php?media=amnis_nome.png"> |
| <table style="border:0px solid white;"> | <table style="border:0px solid white;"> | ||
| <tr style="border:0px solid white;"> | <tr style="border:0px solid white;"> | ||
| <td style="border:0px solid white;"> | <td style="border:0px solid white;"> | ||
| - | <a href="/facility/flowcytometry/lib/exe/fetch.php?media=imagestreamx_wiki.jpg"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=fortessa_1_wiki.jpg" width=300></a> | + | <a href="/facility/flowcytometry/lib/exe/fetch.php?media=imagestreamx_wiki.png"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=imagestreamx_wiki.png" width=300></a> |
| <td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
| <b>Location</b>: <a href="/facility/flowcytometry/lib/exe/fetch.php?media=flow_cytometry_map.jpg">Room P2-A-49</a> (<img src="/facility/flowcytometry/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47224) <br> | <b>Location</b>: <a href="/facility/flowcytometry/lib/exe/fetch.php?media=flow_cytometry_map.jpg">Room P2-A-49</a> (<img src="/facility/flowcytometry/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47224) <br> | ||
| - | <b>Manufacturer</b>: <a href="http://www.bdbiosciences.com/eu/home" target="_blank">BD Biosciences</a><br> | + | <b>Manufacturer</b>: <a href="https://cytekbio.com/" target="_blank">Cytek</a><br> |
| - | <b>Model</b>: <a href="http://www.bdbiosciences.com/us/instruments/research/cell-analyzers/bd-lsrfortessa/m/751752/overview" target="_blank">LSRFortessa</a><br> | + | <b>Model</b>: <a href="https://cytekbio.com/pages/imagestream" target="_blank">Amnis ImageStreamX Mark II</a><br> |
| - | <b>Nickname</b>: "Fortessa 1"<br> | + | <b>Nickname</b>: "ImageStream"<br> |
| - | <b>Software</b>: FACSDiva 6.2<br> | + | <b>Software</b>: INSPIRE 200.1.620.0 / IDEAS 6.2.64.0<br> |
| - | <b>Year</b>: 2010 <br> | + | <b>Year</b>: 2017 <br> |
| - | <b>SN</b>: H5B300013 <br> | + | <b>SN</b>: ISX540 <br> |
|   <br> |   <br> | ||
|   <br> |   <br> | ||
| - | → <a href="https://imm.medicina.ulisboa.pt/files/1414/8700/1713/FORM.UCF.043_-_BD_LSRFortessa_1_and_2_operation.pdf"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=info.gif" width=16> BD LSRFortessa 1 User Guidelines</a><br> | + |   <br> |
| - | → <a href="https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=form.ucf.047_-_bd_lsrfortessa_troubleshooting.pdf"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=info.gif" width=16> BD LSRFortessa Troubleshooting</a><br> | + | → <a href="https://my.agendoscience.com/calendar/?id=MTBjYWxlbmRhcnM=" target="_blank"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=booking.png"> Cytek Amnis ImageStreamX Booking</a> |
| - | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/72"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=date.png"> BD LSRFortessa 1 Booking</a> | + | <br> |
| + | → <a href="/facility/flowcytometry/doku.php?id=amnis_imagestreamx_usage"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=statistics.png"> Cytek Amnis ImageStreamX Usage Statistics</a> | ||
| <br> | <br> | ||
| - | → <a href="/facility/flowcytometry/doku.php?id=bd_lsrfortessa_1_usage"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=chart_line.png"> BD LSRFortessa 1 Usage Statistics</a> | + | → <a href="https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=amnis_brochure.pdf"><img src="/facility/flowcytometry/lib/exe/fetch.php?media=brochure.png"> Cytek Amnis ImageStream Brochure</a> |
|   </p> |   </p> | ||
| </td> | </td> | ||
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| </table> | </table> | ||
| </html> | </html> | ||
| - | {{:info.gif|}} **Next Scheduled Maintenance**: October, 2018 | ||
| ===== System overview ===== | ===== System overview ===== | ||
| - | {{ :cell_analyzers.png?0x300|}} The BD LSRFortessa 1 //cell analyzer// is a benchtop digital flow cytometer equipped with a fixed-alignment flow cell that is gel-coupled to the collection optics. It has three spatially separated lasers - 405 nm, 488 nm and 640 nm. Its primary function is to characterize complex populations of cells. Individual cells pass single file through a stream, which is interrogated by the three lasers with the emitted light being detected by photomultiplier (PMT) tubes. The system electronics calculates the time it takes a single cell to pass through each laser intercept and assigns the signal of each PMT to the appropriate cell. Besides fluorescence, the physical properties of the cell are also analyzed with side scatter (SSC), which reflects the internal complexity of a cell and forward scatter (FSC), which is roughly proportional to cell size. The sample flow rate can be set to low, medium and high values, which correspond approximately to 12, 35 and 60 μl/minute, respectively. If your cells are stained with PerCP-Cy5.5 or you need a high-throughput sampler, check the [[bd_lsr_fortessa_2|BD LSRFortessa 2]] instead. If you need multicolor analysis of up to 16 parameters, check the [[bd_lsr_fortessa_x20|BD LSRFortessa X-20]]. | + | {{ :how_the_imagestream_works.png?0x300|}} The Cytek Amnis ImageStreamX is a benchtop multispectral //imaging flow cytometer// equipped with two CCD cameras that can acquire up to 12 images simultaneously of each cell or object in brightfield, scatter and multiple fluorescent channels at rates up to 5000 objects/sec. It has five lasers - 375 nm, 488 nm, 561 nm, 642 nm and 785 nm (dedicated to side scatter). Cells in suspension are hydrodynamically focused in a flow cell, where they are illuminated by a brightfield light source and by lasers. High numerical aperture (NA) objective lenses collect fluorescence emission, as well as scattered and transmitted light from the cells which then intersect a spectral decomposition element that focus different spectral bands on different physical locations of two CCD cameras. By collecting large numbers of digital images per sample and providing numerical representation of image-based features, the ImageStreamX combines single cell standard microscopy information with the statistical significance of large sample sizes in standard flow cytometry. While you can use it to measure fluorescence intensity as in conventional flow cytometers, the best applications for the ImageStreamX take advantage of its imaging abilities to locate and quantify the distribution of signals within cells or even between cells in cell conjugates. The system also has a dedicated software application used for spectral compensation, image analysis and statistical analysis of acquired images (IDEAS) with predefined wizards for apoptosis, cell cycle, co-localization, internalization, shape change, spot counting and others. |
| - | * Multicolor analysis of up to 10 parameters | + | * **12 imaging channels** (2 brightfield channels + 10 channels for fluorescence detection) |
| - | * Three lasers with violet (405 nm), blue (488 nm) and red (640 nm) | + | * **5 lasers** with UV (375 nm), blue (488 nm), yellow-green (561 nm), red (642 nm) and far red (785 nm) for side scatter |
| + | * **Multimag**: 20x, 40x and 60x magnification | ||
| + | * **Extended depth of field**: projects all structures within a cell into a single plane of focus | ||
| + | |||
| + | {{ ::img-12channels.jpg |}} | ||
| ===== Configuration ===== | ===== Configuration ===== | ||
| - | {{:bd_lsrfortessa_1_configuration_wiki.png?800|}} | + | {{::imagestreamx_fluorochrome_chart.png?700|}} |
| + | |||
| + | ===== Objectives Information ===== | ||
| + | |||
| + | ^ Magnification ^ 20x ^ 40x ^ 60x ^ | ||
| + | ^ Numerical Aperture | 0.50 | 0.75 | 0.90 | | ||
| + | ^ Field of View (μm) | 120 | 60 | 40 | | ||
| + | ^ Pixel Size (μm) | 1.0 | 0.5 | 0.33 | | ||
| + | ^ Depth of Field (μm) | 8.0 | 4.0 | 2.5 | | ||
| + | |||
| + | ===== Image Acquisition Performance ===== | ||
| + | |||
| + | {{::high_speed_imaging.png?600|}} | ||
| + | |||
| + | ===== Additional information ===== | ||
| + | |||
| + | * **[[prepare_samples|How to prepare samples for Flow Cytometry]]** | ||
| + | * [[https://imm.medicina.ulisboa.pt/facility/flowcytometry/lib/exe/fetch.php?media=sop.ucf.026_-_amnis_imagestreamx_operation.pdf| Cytek Amnis ImageStreamX Operation]] | ||
| + | * {{::imagestreamx_sample_preparation_guide.pdf| Sample Preparation Guide}}. **Note**: Samples are run using 1.5 ml microfuge tubes (Sarstedt ref. 72706) | ||
| + | ===== Booking Rules ===== | ||
| + | |||
| + | Available 7 days a week, 24 hours a day for trained researchers. | ||
| + | |||
| + | Please check the {{ ::sop.ucf.004_-_ucf_imm_user_guidelines.pdf |User Guidelines}} for detailed information. | ||
| + | |||
| + | ---- | ||
| + | |||
| + | [[resources|Back to the Resources page]] | ||
imagestreamx.txt · Last modified: 2024/01/22 18:42 by flowcytometry
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