Protocol

Induction of Conjugation in Tetrahymena

This protocol was adapted from "Culture and Manipulation of Tetrahymena," Chapter 18, in Cells (eds. Spector et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA. 1998. This 3-volume set is now out of print; however, some of the microscopy methods were republished in Basic Methods in Microscopy, by Spector and Goldman.

INTRODUCTION

The method to induce Tetrahymena conjugation described below is modified from that of Martindale et al. (1982). Mature Tetrahymena cells of opposite mating types are starved under appropriate salt conditions. The mating types are then combined to costimulate through cell-cell interaction. Loose pairs and then firm, irreversible pairs of cells of opposite mating types form. This method consistently results in a high percentage of pairing (usually greater than 80%) and good synchrony.