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DIVE Multiphoton Microscope Image Gallery

Tunable Multicolor Deep In Vivo Imaging

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Today’s life science research focusses on complex biological processes, such as the causes of cancer and other human diseases. A deep look into tissues and living specimens is vital to understanding the conditions and mechanisms in cells and finding answers to crucial questions challenging the life sciences.

Transcription activity in mice gastric stem cell derived organoid

Hoechst channel labelling the nucleus. The differences in fluorescence lifetime corresponds to difference in DNA compaction. The longer lifetimes (red) correspond to cells with more open reading frames and an active DNA transcription such as stem cells. Video acquired with multiphoton (DIVE) and lifetime (FALCON).

Deep in vivo multicolor imaging

Small intestines of a confetti mouse, fluorescently labeled and lineage traced from a multi-color tracer. The gray color codes for the SHG signal from collagen. Lineage traced stem cells are shown in cyan (CFP), green (GFP), yellow (YFP), and red (RFP). Image size is approximately 700x700x150 μm³. Recorded with two-photon excitation using the SP8 DIVE. Sample courtesy of J. van Rheenen, Netherlands Cancer Institute, Amsterdam (the Netherlands).
Live Mouse Brain cortex. Images acquired on a multiphoton DIVE microscope. Astrocytes labeled with Sulforhodamine (blue); Microglia labeled with GFP (green), Neurons labeled with YFP (yellow) and bood vessels labeled with Alexa680-Dextran (red). Sample courtesy of LMF, DZNE, Germany

Label-free imaging: Second (SHG) and third harmonic generation (THG)

The small intestine of Lgr5CreERT2-GFP confetti mice. Gray: SHG signal (collagen 1) which is used as an orientation landmark. Green: Lgr5+ stem cells. Red, cyan, and yellow, traced cells that are progeny of the Lgr5 stem cells. Sample courtesy of Prof. Jacco van Rheenen, Netherlands Cancer Institute, Amsterdam (the Netherlands).
Mouse brain cortex. Thy1-eYFP cortical neurons, THG blood cells. Excitation: 1300 nm. Objective: IRAPO 25x1.0 W motCorr. 70 fps, 256x256 px. Sample courtesy of Kevin Keppler, Light Microscopy Center, DZNE, Bonn, Germany.

Spectral unmixing

Effortless separation of GFP (green) and YFP (red) expressed in the cortex of a live mouse by capturing either two channels simultaneously or conducting a spectral emission scan and applying channel unmixing in the LAS X software. Sample courtesy of Kevin Keppler, Light Microscope Facility, DZNE, Bonn (Germany).

Three photon excitation

The brain cortex of a live mouse. Thy1-eYFP is expressed in the cortical neurons. Imaged at 450 μm depth in the cortex with 1300 nm excitation wavelength. Sample courtesy of Kevin Keppler, Light Microscope Facility, DZNE, Bonn (Germany).

Deep in vivo colocalization

Striated muscle from a mouse tibia. Morphological analysis of neuromuscular junction development. Green: GFP – Rab 5 (GTPase, endosomes). Red: mCherry – Bif 1. Gray: AF647- Alpha bungarotoxin (NMJ). Sample courtesy of Prof. Rudolf, University of Applied Science, Mannheim (Germany).

Deep in vivo colocalization

Mouse intestine. Blue: Alexa Fluor 350 - wheat germ agglutinin mucus of goblet cells. Green: SYTOX Green - Nuclei. Red: Alexa Fluor 568-Phalloidin - F-Actin, prevalent in the brush border. White: SHG. Sample courtesy of Molecular Probes.

FLIM

Autofluorescence in mammalian cells under non-physiological conditions (pH 8.5). The signal correlates with changes in the NAD/NADH endogenous pool. The development of oxidative stress over time reads out as decrease of fluorescence lifetime. Images size: 512 x 512 pixels. Color scale bar has units of ns. Images are extracts from a movie taken with the SP8 FALCON and SP8 DIVE.

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