This video describes a simple, time-saving technique for automated tissue dissociation using the gentleMACS Dissociator to prepare single-cell suspensions of mouse splenocytes.
Introduction to Equipment
Dissociating the spleen cells
Filtration step
Spleen dissociation using the gentleMACS Dissociator will yield a high percentage of viable splenocytes, which is demonstrated by flow cytometry using the MACSQuant Analyzer. Dead cells are stained by propidium iodide. In the example provided, the forward side scatter dot plot shows the prepared splenocytes.
To avoid tedious and often painful manual dissociations, the gentleMACS Dissociator allows one to dissociate tissue very efficiently under controlled and reproducible conditions. The tissue dissociation procedure is gentle to cells and yields cell suspensions with a high viability rate. The gentleMACS Dissociator in combination with C Tubes is designed for the preparation of single-cell suspensions from various tissues. The instrument is equipped with a number of pre-set programs to dissociate mouse spleen, liver, lung, or neural tissue. The range of programs and protocols will be continuously expanded to include additional tissues.
The authors have nothing to disclose.
Material Name | Typ | Company | Catalogue Number | Comment |
---|---|---|---|---|
gentleMACS™ Starting Kit | Miltenyi Biotec | 130-093-235 | http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx | |
C Tubes | Miltenyi Biotec | 130-093-237 | http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx | |
13 mL Tubes | Miltenyi Biotec | 130-091-596 | http://www.miltenyibiotec.com/en/PG_34_751_13_mL_Tubes_for_LS_Columns.aspx | |
autoMACS™ Rinsing Solution | Miltenyi Biotec | 130-091-222 | http://www.miltenyibiotec.com/en/PG_134_155_autoMACS_Rinsing_Solution.aspx | |
MACS® BSA Stock Solution | Miltenyi Biotec | 130-091-376 | http://www.miltenyibiotec.com/en/PG_33_295_MACS_BSA_Stock_Solution.aspx | |
Pre-Separation Filters | Miltenyi Biotec | 130-041-407 | http://www.miltenyibiotec.com/en/PG_34_247_Pre_Separation_Filters.aspx | |
MACSQuant Analyzer | Miltenyi Biotec | 130-092-197 | ||
Propidium Iodide Solution | Miltenyi Biotec | 130-093-233 | http://www.miltenyibiotec.com/en/PG_337_744_Propidium_Iodide_Solution.aspx |