DCS-120 Super MPC FLIM System

Selected Specifications

Principle

Narrow-band tunable excitation from UV to NIR, fast galvo-mirror laser-scanning, de-scanned confocal detection (DC, confocal mode), non-descanned detection (NDD, multiphoton mode), and bh's multi-dimensional TCSPC FLIM technique

Excitation

• Multiphoton mode: tuneable Ti:Sa fs pulsed excitation, selective excitation in the range from 680 nm to 1300 nm
• Confocal mode: narrow band selective frequency doubling of the multiphoton laser to cover the UV and VIS range of 400 nm – 640 nm. Picosecond duration laser pulses fiber delivered to the microscope for best confocal performance.

Scan Rate, Pixel dwell Time

Down to approx. 1 μs/pixel

General Operation Modes

TCSPC FLIM:

• 2 (multi-) spectral or polarisation channels
• Time-series, Z-Stack, mosaic (x,y, z, temporal)
• FLITS (fluorescence lifetime-transient scanning)
• PLIM (phosphorescence lifetime imaging) simultaneously with FLIM,
• Photon correlation: FCS, FCCS, gated FCS
• single-point fluorescence and phosphorescence decay

Scan Head

Optical Principle

Fast galvo-mirror laser-scanning

Laser Inputs

Two independent inputs, fiber coupled (UV + VIS) and free-beam (NIR)

Optical Laser Power Control

Continuous ND filter wheel control

Confocal Detection

Outputs to Detectors

Two confocal detection detectors directly attached tot he scan head.

Main Beamsplitter Versions

Alignment-free exchangeable dichroics: Longpass, multi-band, wideband, and multiphoton options available

Secondary Beamsplitter Wheel

Three dichroic beamsplitters, polarising beamsplitter, 100% to channel 1, 100% to channel 2

Pinholes

Independent pinhole wheel for each channel

Pinhole Alignment

Electronical, via piezo microstage

Pinhole Size

11 pinholes, from about 0.5 to 10 AU

Emission Filters

Two filter sliders per channel in series

Multiphoton Detection

Non-Descanned (NDD) Detectors

Attached to rear epifluorescence port of the microscope, dichroics and bandpass filters inserted in microscope-standard filter cubes.

NDD Detector Light protection

Fast optical shutte with light level overload protection.

Scan Control

Principle

Hardware controlled precision laser-scanning with fast flyback for rapid acquisition

Frame Size

Frame scan 16 x 16 to 4096 x 4096 pixels, line scan 16 to 4096 pixels

X Scan

Continuous or pixel-by-pixel,

Y Scan

Line-by-line

Electrical Laser Power Control

Software control of a laser with analogue modulation input

Beam Blanking

During flyback and when scan is stopped. Requires software control of laser power.

Frame Rate / Scan Speed

Automatic selection of fastest rate or manual selection

Scan Area Definition

Interactive scan region selection, hardware zoom + offsets.

Fast Preview Function

Yes

Beam Park Function

Yes, interactive measurement point selection.

TCSPC System

TCSPC / FLIM Modules

SPC-180NX

SPC-QC-104

Number of Parallel TCSPC / FLIM Channels

Typ. 2, max. 4

Typ. 2, max. 3

Electrical Time Resolution

1.6 ps RMS / 3.5 ps FWHM

16 ps RMS / <39 ps FWHM

Timing Precision /

1.1 ps

11 ps

Minimum Time Channel Width

405 fs

4 ps

Saturated Count Rate

12 MHz

40 MHz, shared among active channels.

Recording of Multi-Wavelength Data

Simultaneous in 16 channels, via routing function

Operation Modes of TCSPC System

• Hardware pre-analysed imaging
• Photon event stream (FIFO) imaging
• Point measurements for correlation, long timescale intensity (MCS)
• Mosaic imaging, time-series imaging multi-detector operation, cycle and repeat function, autosave function

Software

Data Acquisition Software

bh SPCM, bh LabVIEW for integration of external devices

Scanner Control Software

Integrated in SPCM, bh LabVIEW for integration of external devices

Operation System

Windows 10 / 11 64 bit

Data Analysis Software

bh SPCImage NG

Principle of Data Analysis

MLE fit (GPU assisted processing)

Model of Functions

• IRF convoluted single, double, or triple component exponential decay
• Optional consideration of incomplete decay
• Shifted component model

IRF Modelling

Synthetic IRF function fit to decay data, auto-extraction of IRF from data, or measured IRF

Excitation Sources

Confocal FLIM

Narrow band selective frequency doubling of the multiphoton laser to cover the UV and VIS range of 400 nm – 640 nm. Picosecond duration laser pulses fiber delivered to the microscope for best confocal performance

Multiphoton FLIM

Tuneable Ti:Sa fs-pulsed excitation, selective excitation in the range from 680 nm to 1300 nm.

Detectors

Confocal Detectors

Coupled directly to scan head

NDD Detectors

NDD detectors, coupled directly to back port of microscope. Detector attachment assembly includes space for laser blocking, dichroic and bandpass filters.

Standard Detector

HPM-100-40 hybrid detector with GaAsP cathode, 250 to 720 nm, best for use with ns lifetime dyes

Optional

HPM-100-06 detector with <20 ps FWHM IRF width, 220 to 650 nm, best for ps lifetime autofluorescence studies, confocal detection only.

Optional

HPM-100-50 detector, 400 to 900 nm, best for long wavelength fluorescence

Optional

MW-FLIM GaAsP multiwavelength detector