Culture, Storage and Uses of Physarum Plasmodia, Microplasmodia and Amoeboflagellates

by

Mark R. Adelman


Many of these protocols are simply taken from my lab 'how to' notebooks. They were never meant to be 'formal' publications. Their inclusion here is simply intended to help anyone trying to reproduce and/or extend studies I reported at Physarum meetings or published elsewhere.

PLASMODIA & MICROPLASMODIA

Culture of Microplasmodia - including how to start microplasmodial cultures, how to prepare spherules, and the composition of growth medium.

Culture of Plasmodia - how to start plasmodial cultures in several different ways, how to grow them inexpensively and over a range of culture sizes, and how to harvest plasmodia suitable for biochemical studies.


AMOEBA & AMOEBOFLAGELLATES

BACTERIA - how to grow bacteria to be used as the food source for cultures of amoebae.

Buffers - recipes for various solutions needed to culture amoebae and to fix amoeboflagellates to be used in fluorescence microscopy studies.

Agar Plates - information on how we prepare the agar plates on which Physarum amebae are grown, including the special steps that are taken so amoebae optimized for AFT/RAFT experiments are obtained.

RAFT Cells - definition of the term RAFT and a detailed protocol for the sequence of steps used to prepare such cells for various analyses.

Processing of Amoeboflagellates for Fluorescence Microscopy - discussion of critical steps in the processing, plus a detailed exemplar recipe of how such processing is done.

Shipping Fixed Cells - a simple protocol for preparing fixed cells that you then ship to me for analysis. This is also an open-ended request that you send me such fixed cell samples!