\ iviA.title. 



I BIOLOGiCAl 

 I L/^.BORATORY 



I LIBRARY 



CHAPTER 21 I ^QQQ.. ,,QL£^ ^^^r^^ 



The Cytoplasm } ^■J;^-^^;^" 



GEORGE H. HOGEBOOM and WALTER C. SCHNEIDER 



Page 

 I. Introduction 199 



1. General 199 



2. Cytology of the Cytoplasm 200 



II. Cytochemical Methods 202 



1. General 202 



2. Histochemical Methods 202 



3. Submicromethods 204 



4. The Cell Fractionation Technic 205 



a. Advantages and Limitations 205 



b. Requirements Defining an Adequate Isolation Procedure 206 



c. Method of Cell Disruption 209 



d. The Effect of Various Media on Cytoplasmic Particles and the Cyto- 

 logical Identification of the Components of Cell Fractions 210 



e. Isolation of Mitochondria and Microsomes from Liver 212 



(1) Preliminary Steps 212 



(2) Removal of Nuclei and Intact Liver Cells 212 



(3) Isolation of Mitochondria 212 



(4) Isolation of Microsomes 213 



f. Isolation of Pigment Granules 214 



g. Isolation of Secretory Granules 215 



h. Isolation of Golgi Apparatus 215 



III. Biochemical Properties of Isolated Cell Structures 216 



1. The Nuclear Fraction 216 



2. The Mitochondrial Fraction 217 



a. Biochemical Data Relating to the Integrity of Isolated Mitochondria. 217 



b. Biochemical Properties of Isolated Liver Mitochondria 219 



c. Biochemical Properties of the Mitochondria of Tissues Other than 

 Liver 228 



d. Biochemical Investigations Relating to the Structural Organization 



of Mitochondria 230 



3. The Microsomal Fraction 236 



4. The Soluble Fraction 240 



5. On the Biochemical Homogeneity of Isolated Cytoplasmic Structures . 241 



6. The Pentose Nucleic Acid of Isolated Cell Fractions 244 



I. Introduction 

 1. General 

 Although for many years cytologists have been aware of the great com- 

 plexity of the structural organization of the cell cytoplasm, concrete in- 



199 



