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• W2024717139 abstract "To the Editor: We have a great interest in the recent article by Hayes et al. (1). Because leukocytes act as a major role for the tissue/organ damages, there has been much focus on in vivo visualization of these circulatory leukocytes. Intravital fluorescent microscopy has been established as a versatile technique for the study of cell-cell interaction and blood flow at the level of the microcirculatory unit. While Hayes et al. (1) did not mention how they labeled and detected circulating leukocytes, fluorescent markers such as acridine orange and rhodamine 6G are usually used for direct visualization of leukocytes during these processes (2). However, there are some pitfalls of these labeling methodologies. It has been mentioned that these fluorescent dyes often induce phototoxic effects and arteriolar vasospasm at high concentration levels (2). These dyes are taken up predominantly by leukocytes, but their fluorescent intensity is transient and quickly taken up by hepatocytes 10–20 minutes after IV injection (data not shown). Therefore, this labeling technique does not permit stable identification of individual cell types during observation. To address this issue, we have established a new method for in vivo visualization of leukocyte trafficking using our ubiquitously expressed GFP transgenic (Tg) rats (3). Characteristically, this Tg rat has two peaks of strong GFP intensity in the mononuclear cells and GFP positive granulocytes. Almost all of the granulocytes are detected separately from lymphocytes according to their size and fluorescent intensities (4). Thus, using fluorescent microscopy and charge-coupled-device (CCD) camera, we could visualize the trafficking of GFP-positive leukocytes in the lung and liver in vivo (Fig. 1) (see supplemental Figure 1 as a video file available at anesthesia-analgesia.org).Figure 1.: Successful visualization of GFP positive leukocytes in the liver of a GFP Tg rat using fluorescent microscopy with CCD camera. Arrowhead = leukocytes with bright GFP.Considering the results of our study, we would like to ask how the authors elegantly visualized the activated leukocytes to monitor their dynamics in postcapillary venules using the epifluorescence microscope (1). Aki Sato, MD*†§ Yuki Sato, MD*†§ Yukio Sato, MD, PhD‡ Yasunori Sohara, MD, PhD‡ Norimasa Seo, MD, PhD† Eiji Kobayashi, MD, PhD§ *The first two authors contributed equally to this work. †Department of Anesthesiology ‡ Division of Thoracic Surgery, Department of Surgery §Division of Organ Replacement Research Animal Resource Project Center for Molecular Medicine Jichi Medical School Tochigi, Japan [email protected]" @default.
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• W2024717139 date "2005-04-01" @default.
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• W2024717139 title "Leukocytes with Bright Fluorescence in Rats" @default.
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• W2024717139 doi "https://doi.org/10.1213/01.ane.0000149018.17929.9d" @default.
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