Table of Contents
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Location: [Oeiras] Room 0B10 |
Miltenyi UltraMicroscope Blaze Quality Control
Miltenyi UltraMicroscope Blaze Usage StatisticsMicroscope overview
The UltraMicroscope Blaze is a lightsheet for large cleared samples system with dual sided illumination, capable of using three beams per side. This allows for uniform imaging with minimal shadows (in good sample clearing conditions). Equipped with three objectives (1.1x, 4x and 12x) and internal magnifications of 0.6x, 1x, 1.66x and 2.5x, this system is capable of going from whole organs to single cell level. There are 4 laser lines in the Blaze, 488, 561, 640 and 785 nm, and with the capability of combining these with any of the installed filters, the system is capable of mix-matching for markers such as Propidium Iodide.
Usage conditions
The default configuration for the UltraMicroscope Blaze is for the Refractive Index of DBE (iDisco protocol) at RI = 1.56. Any use where another immersion media is necessary, the system will need realignment, and possibly undergo objective dipping-cap changes (below RI = 1.49). The time taken to do these alignments (DBE to immersion media, immersion media to DBE) can vary between 1-3 hours, and will be charged to the user in question.
Laser Lines
| Wavelength (nm) | Fluorophores | Penetration capability |
|---|---|---|
| 488 | E.g: GFP, TRITC, AL488 | Less penetration |
| 561 | E.g: mCherry, RFP, AL594 | Good penetration |
| 640 | E.g: Cy5, TexasRed, AL647 | High penetration |
| 785 | E.g: Atto740, Atto790 | Highest penetration |
Fluorescence filters
| Position | Filterset | Emission (nm) |
|---|---|---|
| 1 | 488 | 525/50 |
| 2 | 561 | 595/40 |
| 3 | 640 | 680/30 |
| 4 | 785 | LP 805 |
| 5 | Empty | - |
| 6 | Empty | - |
| 7 | Empty | - |
Objectives
| Magnification | Dipping caps RI range | NA | WD per dipping cap (mm) | Maximum Theoretical Resolution (um) |
|---|---|---|---|---|
| 1.1x | 1.33-1.49 / 1.50-1.57 | 0.1 | 16 / 17.6 | 4.8 |
| 4x | 1.33-1.41 / 1.42-1.48 / 1.49-1.57 | 0.35 | >=15 / >=15 / >=15 | 1.3 |
| 12x | 1.33-1.41 / 1.42-1.48 / 1.49-1.57 | 0.55 | 8.5 / 10.9 / 10 | 0.5 |
Objectives + Zoom
| Objective | Zoom | Final Magnification | Field of view (mm) |
|---|---|---|---|
| 1.1x | 0.6x | 0.66x | |
| 1.1x | 1x | 1.1x | |
| 1.1x | 1.66x | ~1.83x | |
| 1.1x | 2.5x | 2.75x | |
| 4x | 0.6x | 2.4x | |
| 4x | 1x | 4x | |
| 4x | 1.66x | 6.64x | |
| 4x | 2.5x | 10x | |
| 12x | 0.6x | 7.2x | |
| 12x | 1x | 12x | |
| 12x | 1.66x | 19.92x | |
| 12x | 2.5x | 30x |
Camera
| Model | Frame Size | Pixel Size (µm) | Quantum Efficiency |
|---|---|---|---|
| PCO Edge 5.5 sCMOS | 2048 × 2048 | 6.5 x 6.5 | 60% |
Microscope Turn On Procedure
- Turn on the power bar on top of the microscope's computer, under the table.
Microscope Turn Off Procedure
- With the software still open, pull out the cuvette drawer.
- Wait for the drawer to move all the way down.
- Remove any used cuvette.
- Press the drawer button, but do not push the drawer in.
- In software, change to 12x objective and lower the objectives all the way.
- Turn off the power bar on top of the microscope's computer, under the table.
- Close the cuvette drawer and software.