粒線體螢光染劑-適用活細胞

發布日期:2024/7/31

震撼登場 !

研究人員已擁有先進的顯微影像系統, 諸如超解析奈米影像系統, 共軛焦影像系統, 及各類型顯微鏡影像系統, 總是希望取得更高解析更精細的細胞結構與動態交互關係, 要達到此一目的, 需要新一代的螢光染劑 !


PK Mito 粒線體螢光染劑就是最新的突破, 全新的設計, 其光毒性降到最低, 可以提供更長時間的活細胞實驗, 取得最高解析的影像品質.








新一代超高解析的活細胞或固定細胞適用的 PK Mito Orange 粒線體螢光染劑, 使用簡單,無需轉染,僅需15分鐘染色,便可得到絕佳的超解析影像, 獲得穩定的高亮度螢光信號,並能夠在成像時最大程度保持粒線體的自然生理狀態,使觀察活細胞時的光毒性干擾降至最低,並顯著延長對線粒體的觀察時間。是絕佳的超解析及長時間粒線體成像染料.


PK Mito - ( 俗稱 PKMO ) - 專屬應用於活細胞的螢光染劑, 目前有 PK Mito Orange, PK Mito RED, PK Mito Deep Red 三款 粒線體染劑




PKMO in conjunction with fluorogenic rhodamine probes enables nanoscopic mapping of mitochondria and the analysis of mito-organelle interactions using multi-color STED nanoscopy. A. Labeling strategies for multi-color live-cell recordings of mitochondria. Cartoon demonstrating the labeling strategies for mitochondrial subcompartments and for mitochondria-interacting cellular structures. Abbreviations: ER (endoplasmic reticulum); IM (inner mitochondrial membrane); OM (outer mitochondrial membrane); CJs (crista junctions), mtDNA (mitochondrial DNA). B–G. Multi-color live-cell STED nanoscopy of HeLa and COS-7 cells labeled for the IM together with different mitochondrial targets or subcellular structures. IM was labeled using PKMO and recorded by live-cell STED nanoscopy (λex = 561 nm, λSTED = 775 nm). B. 2D single-color STED nanoscopy of mitochondria labeled for mtDNA. The mtDNA was stained using PicoGreen and was recorded in the confocal mode (λex = 488 nm). C and D. Dual-color STED nanoscopy of mitochondria in COS-7 cells. OM marker TOM20-Halo was labeled using 647-SiR-CA (λex = 640 nm, λSTED = 775 nm). Cells were recorded by 2D STED in (C) or 3D STED nanoscopy (D). E. 2D dual-color time-lapse STED nanoscopy of mitochondria in HeLa cells. CJs were labeled by overexpression of MIC10-SNAP and staining with SNAP-Cell 647-SiR. Insets (Lower panel) shows four consecutive frames illustrating cristae and CJ dynamics. F. 2D dual-color STED nanoscopy of mitochondria and microtubules in HeLa cells. Microtubules were labeled using 4-610CP-CTX (λex = 640 nm, λSTED = 775 nm). MtDNA was labeled using PicoGreen. Inset highlights contact sites of mitochondria and microtubules. G. 2D dual-color time-lapse STED nanoscopy of mitochondria and ER in HeLa cells. ER was labeled by overexpression of Halo-KDEL and staining with 647-SiR-CA. Insets (Lower panel) highlights contact sites of a mitochondrion and ER over several time points. If not indicated otherwise, all image data were deconvoluted and corrected for photobleaching. (Scale bars, 2 µm.)



參考文獻:
 
1. Yang, Zhongtian, et al. "Cyclooctatetraene-conjugated cyanine mitochondrial probes minimize phototoxicity in fluorescence and nanoscopic imaging." Chemical Science (2020).
 
2. Liu, Tianyan, et al. "Multi-color live-cell STED nanoscopy of mitochondria with a gentle inner membrane stain." Proceedings of the National Academy of Sciences 119.52 (2022): e2215799119.



 


PK Mito Orange Fix 適用於固定細胞的粒線體染劑

PK Mito Orange Fix 粒線體探針 是用於固定樣本的 粒線體內膜螢光染料,能夠特異性結合到各類細胞的粒線體內膜上,繼而利用醛類固定液固定後成像。與傳統染料相比,PKMITO® 系列 粒線體染料 光毒性大幅度降低,使用更加簡便,只需染色30分鐘後按普通方式固定,就能獲得穩定的高亮度螢光信號,並能在成像時最大程度保持粒線體狀態,是絕佳的超解析及長時間粒線體成像染料. ( 本染劑, 也有名稱為 PK Mito Orange FX )

本產品在固定後可達到高於 100nm 的成像解析度,並向下相容各種螢光顯微鏡。

每份PKMITO®Orange Fix 染料足夠 10-50 次成像。







PKMO FX enables nanoscopic STED imaging of mitochondria in fixed cells. ( A) STED imaging of mitochondrial cristae of a fixed COS-7 cell labeled with PKMO FX and fixed with 2.5% GA. (Scale bar, 2 μm.) ( B) STED imaging of mitochondrial cristae of a fixed COS-7 cell labeled with PKMO FX and fixed with 4% FA. (Scale bar, 2 μm.) ( C) STED imaging of mitochondria from a fixed HeLa cell labeled with MTR and fixed with 2.5% GA. (Scale bar, 2 μm.) (D and E) STED imaging of mitochondria labeled with PKMO FX in the same area before and after FA/GA fixation. After live cell imaging, 4% FA (D) or 2% GA (E) fixative was added to cells and STED images were acquired after 3 min. (Scale bar, 2 μm.) (F) Z-stack STED images of a HeLa cell labeled with PKMO FX and fixed with 2% GA. Color code image shows the whole-cell 3D reconstruction from 14 individual z-section images. Step size = 156 nm. (Scale bar, 10 µm.) (G, Left) Magnified images of the white boxed areas in F. (Scale bar, 5 µm.) Right: two close-up z-section images from frame #3 and #10. (Scale bar, 2 µm.)


1. Chen, Jingting, et al. "An aldehyde-crosslinking mitochondrial probe for STED imaging in fixed cells." Proceedings of the National Academy of Sciences 121.19 (2024): e2317703121. 




保   存 :  避光, 冷凍,  攝氏 - 20 度
有效期 : 1 年



 



PK MEM 系列 的細胞膜探針具有極低的化學毒性、光學毒性, 具有優秀的細胞膜定位能力,適用於多種癌細胞和原代細胞的細胞膜標記,可進行活細胞或固定細胞的超解析成像。本系列產品也已經在小鼠活體雙光子實驗中實現神經元細胞膜成像。

PK MEM系列產品適用實驗場景廣泛,性能優異,尤其在共軛焦, 多光子, SIM和STED等超分辨長時間成像和細胞分選等領域表現優秀。

每份 PK MEM 系列產品可以滿足10/50次超解析成像。



Ling, Jing, et al. "A gentle palette of plasma membrane dyes." bioRxiv (2024): 2024-05.