Lab Essentials for Craft Brewing, Distillation and Beverage

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Simple and Cheap Yeast Related Analytical Methods and Techniques

Eric AllainAssociate Professor

Brewing Science ResearchDepartment of Chemistry

Outline

● Yeast Counting Using a Microscope and Hemacytometer

● Microorganism Collection and Storage

Yeast Counting With a Microscope and Hemacytometer

Why?

Yeast Counting With a Microscope and Hemacytometer - What You Need

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What the Heck Is a Hemacytometer?

A Look At A Hemacytometer Through A Microscope

How to Use a Hemacytometer to Determine Yeast Cell Concentration

● Volume of counting area is 1mm x 1mm x 0.1mm = 0.0001 ml

● Cells in counting area / 0.0001 ml = cell concentration in cells/ml

● Total cells = cell concentration x total volume of culture

Dilutions

Example 1:100 Dilution

10 ul yeast cells990 ul water

or1 ml yeast cells99 ml water

Multiply result by 'dilution factor' to get total yeast cell concentration

Note: EDTA for resuspending flocculated yeast

More Microscope Fun

● Live/dead staining with methylene blue

● Looking for bacteria or wild yeast contamination

● Checking out barrel aging cultures

● Adding a cheap camera to record images

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Microorganism Collection and Storage

Why?

Microorganism Collection and Storage - What You Need

Media Preparation

Make a 10 Plato (1.040 SG) wortAdd 20 g/L of agarSterilize with pressure cooker along with petri plates and tubesPour plates and slants before media completely cools

Collecting and Storing Yeast on Plates and Slants

Sterilize inoculating loopDip loop into the cultureStreak loop onto plateStore plates 'upside down' at an appropriate temp to let yeast growUse a similar procedure to inoculate slants for longer term storage

Starting a Culture From a Plate or Slant

Reverse the collection procedureUse (cooled) sterile loop to collect a small dab of yeast from the plate or slantDip the loop into a small (~ 5 ml) tube of sterile wort (10 plato)Grow for 2-3 daysUse to inoculate 50-100 mls of sterile wort and grow for 2-3 daysContinue until desired culture volume is reachedMonitor total cells using hemacytometer