Overview

This microscope is a super resolution microscope based on the Leica SP8 confocal microscope.

This instrument is based around an inverted Leica SP8X white light confocal microscope equipped with 405nm and a pulsed White Light Laser (470-670 nm) sources. This white light laser (WLL) source of the Leica SP8X perfectly matches the wavelenght of any fluorophore. Up to eight excitation lines can be used simultaneously. By tuning both excitation and detection, complete excitation-emission spectra can be acquired.With this spectral information, any dye can be optimally excited with minimum cross-excitation and specimen damage. 

In addition, the pulsed WLL supplies excitation light for fluorescence lifetime measurements. Each selected wavelength has about 1.5 mW power which can be attenuated with the AOTF (Acusto-Optic Tuneable filters). The WLL is a pulsed laser (80 MHz), allowing time gating of fluorescence life times in combination with Hybrid detectors. 

The system has two PMTs for fluorescence, one for transmitted light and two gated hybrid detectors. Hybrid detectors allow Low Dark Noise and High Contrast: Sensitive live specimens need to be imaged under low light conditions in high gain situations. Low noise can become the decisive advantage when it comes to recognizing weakly stained detail. Low dark noise is necessary for maximum signal efficiency, especially when photons are accumulated for more information. Otherwise, noise will pile up in the background of the image. 

The Leica HyD offers superior signal-to-noise ratio to help render the finest details from any specimen – even tricky ones, such as highly scattering tissue slices. By reducing dark noise, the Leica HyD automatically improves image contrast. You obtain more information content, and the images are immediately publication-ready without any need for image processing. An incubation box allows temperature and Co2 control. Adaptive focus control (AFC) maintains the focus during prolonged live cell experiments. 

Imaging modes

• Multi point x,y (2D) with tiling,
• Volume x,y,z (3D),
• Time lapse x,y,z,t (4D) 
• DIC
• Live cell
• FRET
• FRAP
• STED superresolution

Specification

Here you can find the specification for this microscope.

Objectives

Magnification	NA	Coverslip	Other modes	Immersion
10	0.04	0.17		Air
20	0.75	0.17	DIC	Air
40	1.3	0.17	DIC	Oil
63	1.2	0.17		Water
63	1.4	0.17		Oil
100	1.4	0.17	STED White	Oil

 

 

 

 

 

 

 

 

 

Filter (epi)

Cube	Example fluorescence	Excitation	Dichroic	Emission	Part number
Leica DAPI	DAPI, Hoechst	340-380	400	420-(LP)
Leica FITC	GFP, FITC, AF488	450-490	510	515 (LP)
Leica RHOD	mCherry, AF594	515=560	580	590 (LP)

Light source (epi)

Broad spectrum fluorescent light

Light source (laser lines)

White Light Laser - any wavelength between 470nm to 670nm

Detectors (cameras or PMTs)

2 HyD, 2 PMT, 1 TL

Location

Leech Building, M2.147a

Medical School
Newcastle University
Newcastle upon Tyne
NE2 4HH

Booking

Find out how to book this microscope.

Booking information

• The minimal booking time for this system is 30 minutes. 
• Time increments of 15 minutes are permitted.
• Bookings can be cancelled with no charge up to an hour before scheduled use

Training video

This tutorial video shows you how to operate the microscope including start-up, image capture, cleaning procedure and shutdown.

SOP

Leica SP8STED SOP