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Cat. No.: 3D7A-100 (for 100T)

Cat. No.: 3D7A-500 (for 500T)

Description

SBS3D™ 7-AAD Cell Viability Assay Kit is developed based on the Far-red fluorescent probe 7-AAD (7-aminoactinomycin D), which specifically stains necrotic cells with compromised membrane integrity in cell spheroids or organoids. This kit allows for rapid, convenient, and specific nuclear staining of necrotic cells in 3D cultured cell spheroids or organoids. With just 10 minutes of staining, bright far-red fluorescence can be observed under a fluorescence microscope. The kit can be used in combination with SBS3D™ Annexin V-FITC/EGFP/PE, Calcein AM, etc., for the detection of apoptosis and necrosis or cell viability in 3D cultures. It is suitable for use with fluorescence microscopes, fluorescence microplate readers, and other fluorescence detection systems.

7-AAD, also known as 7-aminoactinomycin D, is a fluorescent indicator that can intercalate into nucleic acids. The resulting DNA complex produces maximum emission at 647nm when excited at 546nm, making it suitable for multicolor fluorescence microscopy or flow cytometry experiments.

Similar to propidium iodide (PI), 7-AAD is a non-cell membrane permeable dye and cannot pass through the intact cell membrane, allowing it to distinguish between viable and necrotic cells. It can also be used for nucleic acid staining of fixed or permeabilized cells. Unlike PI, 7-AAD has a narrower emission spectrum and longer emission wavelength when excited by a 546nm argon-ion laser, causing less interference with other detection channels, making it an ideal substitute for PI in multicolor fluorescence analysis. It can be used in conjunction with various fluorescent dyes excited by 488nm light, such as FITC (fluorescein isothiocyanate), PE (phycoerythrin), APC (allophycocyanin), Calcein AM, etc. Weak fluorescence is detectable around 600nm by standard fluorescence microscopy, while strong far-red fluorescence around 650nm can be detected by flow cytometry in the FL3 channel or fluorescence microscopy equipped with a 650nm long-pass filter.

The molecular formula of 7-AAD is C62H87N13O16, with a molecular weight of 1270.45, and the CAS number is 7240-37-1. When intercalated into DNA, the maximum excitation wavelength is 546nm, and the maximum emission wavelength is 647nm.

Traditional cell culture is predominantly conducted in a two-dimensional (2D) format. However, cells grown in 2D culture differ significantly from those in vivo in terms of growth patterns, morphology, differentiation, and function. These differences, stemming from the lack of proper cell architecture and tissue morphology, can undermine the reliability of experimental results. In contrast, three-dimensional (3D) cell culture better simulates the microenvironment in which cells exist in vivo, more accurately representing in vivo tissues and reflecting interactions between cells, and between cells and the matrix. The response of cells to exogenous and endogenous stimuli in 3D culture closely mimics their in vivo reactions, making 3D cell culture a more valuable and reliable in vitro experimental model that yields results more consistent with in vivo experiments.

3D tumor cell models are increasingly being used to understand disease mechanisms and in drug development. While 2D cultured tumor cells can uniformly receive nutrients and oxygen from one side, cells within 3D cultured tumor spheroids have less access to these resources, forming natural gradients of nutrients and oxygen. This better simulates the in vivo microenvironment, making 3D cultured tumor spheroids or organoids more effective models of in vivo tumors. They are particularly useful for small molecule drug screening and research into tumor-related molecular mechanisms, and can more accurately predict in vivo drug responses, efficacy, and toxicity. Additionally, 2D tumor cell models face limitations in expansion, often losing the genetic heterogeneity of the original tumor and exhibiting clonal selection, reducing clinical relevance. Compared to 2D cell models, 3D cell spheroids or organoids often provide more reliable research results and streamline drug evaluation processes. Since the first establishment of small intestine organoids in 2009, research on 3D cells and organoids has expanded to many tissue systems and become one of the hottest fields in life sciences.

For 96-well plates requiring 100μl of SBS3D™ 7-AAD Cell Viability Detection Working Solution per well, the small package of this kit can test 100 samples, while the medium package can test 500 samples.

Features

• This kit has a wide range of applications. It can be used for 3D cell spheroids or organoids cultured by conventional methods, including ultra-low attachment plates, Matrix-Gel or Matrigel-coated plates, agarose-coated plates, hanging drop culture plates, etc.
• The kit is easy to use, with the entire detection process taking approximately 10-30 minutes. After treating 3D cell spheroids with apoptosis or necrosis inducers, prepare SBS3D™ 7-AAD Cell Viability Detection Working Solution by diluting SBS3D™ 7-AAD Staining Solution (100X) with Detection Buffer as specified, and incubate in the dark for 10 minutes for subsequent fluorescence microscopy imaging and analysis.

Components

• SBS3D™ 7-AAD Staining Solution (100X)
• Detection Buffer