Moxi Flow
Yeast Count SOP
EtOH Produc,on Materials
Item QTY Picture Item QTY Moxi Flow ORFLO/White Labs Yeast Analyzer Vendor: White Labs PN: MB2000 1 500ml Moxi Flow MFS CasseZe Vendor: Midland PN: MB2000 1 PBS Vendor: GIBCO PN: 10010049 675ul P100 PipeZe (set to 75ul) Various 1 Moxi Cyte Viability Reagent Vendor: Midland PN: MXA054 P1000 PipeZe (set to 675ul) Various 1 1 PipeZe ,ps (for p100 and p1000) Various 1 1.5 ml Eppendorf Tube Various 200 ml Erlenmeyer flask 1 Picture Yeast Prep Procedure
Step Procedure 1 Place empty 200ml Erlenmeyer Flask onto scale and tare to zero 2 Weigh out 98.75 fluid grams of source PBS into the 200ml Erlenmeyer Flask 3 Weigh out 1.25 fluid grams of tank sample into the 200ml Erlenmeyer Flask 4 PipeZe 675ul’s of source PBS into Eppendorf Tube 1 and 675 ul’s of Moxi Cyte Viability reagent into Eppendorf Tube 2 Notes 1 2 Yeast Prep Procedure
Step Procedure 5 PipeZe 75ul’s from THOROUGHLY MIXED 200ml Erlenmeyer Flask into Tube 1 and then pipeZe 75 ul’s from thoroughly mixed Tube 1 into Tube 2, cover with Aluminum foil for 2 minutes 6 Select “Yeast Count” applica,on on Moxi Flow by tapping with finger 7 Insert the casseZe, close the door and system will auto align the laser 8 When Alignment is complete, open the door and pipeZe 75ul’s from thoroughly mixed sample in step 5 into casseZe and close the door Total dilu:on factor =8,000X Mul,ply final result by dilu,on factor to calculate yeast sample concentra,on Notes 1 2 Calculating Total Yeast
Count
Step Procedure 9 Total Yeast Count Press RCL buZon and select “0” Record live (6.21x10^4) cell count in excel template. Note gate is set as part of the instrument installa,on, using the “STO” func,on on the screen. Total % Viability is listed in the yellow box at top of screen (77.9%) Notes Note this is the valley between budding yeast (to the right) and single cell yeast popula,on (to the leg Calculating % Budding
Step Procedure 10 Total Yeast Count Press RCL buZon and select “1” Record live budding cell count (3.98 x 10^4) in excel template. % Budding = 3.98/6.21 = 64% Note gate the leg ver,cal gate is now set at the valley between the single cell yeast popula,on and the budding yeast popula,on Note 2: The “Yeast Count” Applica,on was used in this example. As per the guidance of WhiteLabs, who sets the standards for yeast coun,ng in the brewing markets yeast buds are counted as 2 yeast cells (or 3, 4, etc if chaining) and therefore % budding will be higher than if you tradi,onally count yeast buds as a single cell. Select the “Viability Count (PI) applica,on if you prefer coun,ng yeast buds as a single cell Notes Note this is the valley between budding yeast (to the right) and single cell yeast popula,on (to the leg UWGP Example
Calcula9on Total Yeast Count Press RCL buZon and select “1” Record live budding cell count (3.98 x 10^4) in excel template. % Budding = 3.08/5.25 = 58.7% Note gate the leg ver,cal gate is now set at the valley between the single cell yeast popula,on and the budding yeast popula,on Note 2: The “Yeast Count” Applica,on was used in this example. As per the guidance of WhiteLabs, who sets the standards for yeast coun,ng in the brewing markets yeast buds are counted as 2 yeast cells (or 3, 4, etc if chaining) and therefore % budding will be higher than if you tradi,onally count yeast buds as a single cell. Select the “Viability Count (PI) applica,on if you prefer coun,ng yeast buds as a single cell Notes Note this is the valley between budding yeast (to the right) and single cell yeast popula,on (to the leg