Bioimaging Wiki

Microscope 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
6	Fura-2	370-390 nm	400 nm	498-518 nm

Emission Filters Fast Filter Wheel

Filterset	Emission	Reference
DAPI-FW	435-485 nm	AT460/50m
FITC-FW	500-550 nm	ET525/50m
TRITC-FW	570-620 nm	ET595/50m
Cy5-FW	665-705 nm	S685/40m

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
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
100x	Plan Apo Lambda	Oil	1.45	0.13	Yes	Yes (N2)	No	MRD01905*	0.065	0.043

* 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.

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