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1400 x 765 μm² tile scan of large intestine of an anasthesized mouse. The blood system (black) is visualized using the instrinsic label-free THG signal. Excitation was 1275 nm, non-descanned detection 420-430 nm.
1400 x 765 μm² tile scan of large intestine of an anasthesized mouse. The blood system (black) is visualized using the instrinsic label-free THG signal. Excitation was 1275 nm, non-descanned detection 420-430 nm.
Article

How to Quantify Label-free Blood Flow

The application note “Label-free Blood Flow Quantification” is a guide for data acquisition and analysis of blood flow using label-free third harmonic generation (THG) imaging. Measuring the blood flow rate is very useful for determining the overall status of an organism, organ, or tissue for research in various life science fields like neuroscience, immunology, and cancer. Blood flow tracking is possible with THG due to long IR excitation wavelengths and easy-to-use spectral non-descanned detection. By exploiting the intrinsic contrast of red blood cells, labeling becomes unnecessary. The elimination of the need for labeling not only saves the researcher’s time, but also reduces the risk of image artifacts which can be induced by labels or the labeling procedure.

The described workflow was performed with a state-of-the-art SP8 DIVE multiphoton microscope equipped with a long-IR laser and flexible detection. The workflow did not require any additional hardware or software.

1400 x 765 μm² tile scan of large intestine of an anasthesized mouse. The blood system (black) is visualized using the instrinsic label-free THG signal. Excitation was 1275 nm, non-descanned detection 420-430 nm.

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