Multiphoton Microscope Modules

FLIM & PLIM Module

The dedicated solution for advanced fluorescence and phosphorescence lifetime imaging applications

About Bruker's FLIM/PLIM Module

Many biological processes — including synaptic plasticity — involve dynamic interactions between proteins. Unfortunately, neither classical biochemical approaches nor normal diffraction-limited fluorescence microscopy provides the required sensitivity or resolution to study these intimate and sometimes transient interactions (1).

Contrasting these methods, Fluorescence Lifetime Imaging Microscopy (FLIM) delivers information about changes in the binding, conformation, and composition of biological samples that are not accessible using other techniques. It stands out as the most rigorous method for measuring:

  • Molecular environment parameters;
  • Protein-interactions by Fӧrster resonance energy transfer (FRET) (2); and
  • The metabolic state of cells and tissue via their autofluorescence (3).
High resolution two-photon excitation fluorescence intensity image of mouse epithelium. Intrinsic NADH fluorescence is in green, while autofluorescence from keratin is in red/yellow. Cell morphology can be inferred from NADH signal, which primarily emanates from mitochondria. Data courtesy of Dr. Kyle Quinn, Univ. of Arkansas.
NADH Imaging of developing mammary tumors in vivo (MMTV-PyVT mouse model). Labels: NADH (red), GFP-CFMS (green), SHG (grey). Data courtesy of Dr. Kevin Eliceiri’s lab, Univ. of Wisconsin, Madison. Find more details by clicking on the image.

To do this, FLIM-based FRET measures the change in the decay function of the FRET donor on interaction with an acceptor. Similarly, Autofluorescence FLIM differentiates changes in the biology of objective cells and tissues based on the altered decay parameters of relevant endogenous fluorophores.

Likewise, Phosphorescence Lifetime Imaging Microscopy (PLIM) has become increasingly popular due to the recent development of two-photon-excitable oxygen-sensing probes (4). PLIM allows for quantitative in vivo oxygen measurements at high spatial and temporal resolution with high specificity, which makes it uniquely useful for live-cell and metabolic imaging.

Data courtesy of Dr. Anna Devor’s lab, Boston Univ. Find more details by clicking on the image.
Data courtesy of Dr. Anna Devor’s lab, Boston Univ. Find more details by clicking on the image.

How the FLIM/PLIM Module Supports Advanced Imaging

To leverage these unique capabilities, Bruker has developed a FLIM module based on Becker & Hickl 's FLIM systems. This module is fully integrated with all of Bruker’s multiphoton microscope systems, while Bruker’s proprietary Prairie View software enables advanced FLIM and PLIM data acquisition.