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Table of Contents
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Location: [Oeiras] Room 0B05 |
Nikon HTM-HCS Quality Control
Nikon HTM-HCS Usage StatisticsMicroscope overview
The Nikon HTM-HCS is a fully automated microscope capable of high-throughput imaging and screening of samples (eg, Ibidi chamber slides or well-plates); with the help of fully programmable and customizable acquisition protocols.
The system is equipped with a CoolLED fluorescence illuminator with 4 excitation bands, and an Andor Zyla 4.2 sCMOS camera with 4.0 MPixels with a large field of view capable of maximum of 100 FPS at full frame (2048×2048).
The optics available include fluorescence filters and brightfield, including DIC (for several magnifications) and phase-contrast (40x LWD only).
This microscope is also equipped with a Ionoptix Calcium and Contractility system for high-speed simultaneous acquisition of Fura-2 fluorescence with video-based cell length measurement to study myocyte function, contracting spontaneously or with field stimulation.
- High-speed contractility camera: MyoCam-S3
- Electrical Field Stimulator: MyoPacer
- LED Light Source: UV LEDs for Fura-2 excitation
- High-speed PMT: PMT400 for photon counting at 1kHz
- Software: IonWizard 7.7.3.180
System Components
LEDs
This system is equipped with a CoolLED fluorescence illuminator with 4 excitation lines:
| Line | Wavelength |
|---|---|
| UV | 380 nm |
| B | 470 nm |
| G | 550 nm |
| R | 635 nm |
Filtersets Turret 1 (top)
| Position | Filterset | Excitation | Dichroic | Emission | Reference |
|---|---|---|---|---|---|
| 1 | QUAD | 349-407 nm | 409 nm | 412-452 nm | FL-007110 |
| 459-489 nm | 493 nm | 497-533 nm | |||
| 541-567 nm | 573 nm | 577-613 nm | |||
| 624-646 nm | 652 nm | > 658 nm | |||
| 2 | Green | 460-488 nm | 495 nm | 503-548 nm | |
| 3 | Orange | 532-554 nm | 562 nm | 573-613 nm | |
| 4 | Red | 624-642 nm | 652 nm | 659-701 nm |
Filtersets Turret 2 (bottom)
| Position | Filterset | Excitation | Dichroic | Emission |
|---|---|---|---|---|
| 6 | Fura-2 | 334-346 nm | 400 nm | 498-518 nm |
| 370-390 nm |
Fast Filter Wheel Emission filters
Objectives
| Magnification | Model | Immersion | NA | WD (mm) | PFS | DIC | PhC | Reference | Pixel size (um) (1x zoom) | Pixel size (um) (1.5x zoom) |
|---|---|---|---|---|---|---|---|---|---|---|
| 10x | Plan Apo | Air | 0.45 | 4 | No | Yes (N1) | No | MRD70170 | 0.643 | 0.429 |
| 20x | Plan Apo Lambda | Air | 0.75 | 1 | Yes | Yes (N1) | No | MRD00205* | 0.323 | 0.216 |
| 100x | Plan Apo Lambda | Oil | 1.45 | 0.13 | Yes | Yes (N2) | No | MRD01905* | 0.065 | 0.043 |
| 40x | Plan Fluor | Oil | 1.3 | 0.24 | Yes | Yes (N2) | No | MRH01401 | 0.162 | 0.108 |
| 40x | Plan Apo | Air | 0.95 | 0.17-0.25 | Yes | Yes (N2) | No | MRD00400* | 0.161 | 0.107 |
| 40x | LWD* * | Air | 0.55 | 2.1 | No | No | Yes | MRP46402 | 0.161 | 0.108 |
* Not fabricated anymore ; * * LWD - Long Working Distance
The Bioimaging Platform highly advises against using the 40x LWD objective with 1.5x zoom
Camera
| Model | Frame Size | Pixel Size (µm) | Quantum Efficiency |
|---|---|---|---|
| Andor Zyla 4.2 PLUS | 2048 × 2048 | 6.5 x 6.5 | 82% |
Microscope Turn On Procedure
- Turn on the microscope using the switch on the bottom of the microscope's optical table.
Microscope Turn Off Procedure
- Turn off the microscope using the switch on the bottom of the microscope's optical table.
