THE EXPERIMENTAL COURSE

OF CELL BIOLOGY

Welcome to Study on ECCB2015

Exp3-Staining of the membrane vesicle system
Objectives:
 To grasp the staining method of the membrane
vesicle system using Neutral Red Staining
Solution.
 To observe the basic structure of the
membrane vesicle system, especially the Golgi
complex
 Introduction
 Membrane vesicle trafficking in eukaryotic animal cells
involves movement of important biochemical molecules from
Golgi body to specific 'release' locations of the secretory cell.
 Membrane vesicles (MVs) are termed by the form of Golgi
membrane-bound micro-sized vesicles. MVs packed cellular
products are released/secreted outside the cell across plasma
membrane.
 Vesicular membrane is retained and recycled by the secretory
cells. This phenomenon has key role in synaptic
neurotransmission, endocrine secretion, mucous secretion,
granular-product secretion by neutrophils,etc.
The scientists behind this discovery were awarded Nobel prize for
the year 2013.
The structure of Golgi complex
Functions of Golgi complex: protein transporting and
sorting
 Introduction

 Neutral red is used as a general stain for histology. It stains

lysosomes red. It is used as a counter stain in combination
with other dyes, and for many staining methods. Together
with Janus Green B, it is used to stain embryonal tissues and
supravital staining of blood. Can be used for staining Golgi
apparatus in cells and Nissl granules in neurons.
 Camillo Golgi, using silver staining, identified the
intracellular reticular apparatus in 1898 which bears his
name, the Golgi apparatus.
 Contents

1. Specimen preparation of the Staining and

observation of the membrane vesicle system
using Neutral Red Stain Method.
2. To observe the Golgi apparatus
 Materials and methods
1. Materials
 Frogs (Rana catesbeiana)
 Materials for frog dissection include forceps,
scissors, and teasing needle etc.
 Reagents for sample preparation and staining
include Ringer's solution, and neutral red
staining solution.
 Silver pre-stained slides of neuron .
 Microscopes
2. Methods
 Specimen preparation: open the chess of a frog → find
out the xiphoid bone and take out the thinnest part (about
3-4mm2)→ rinse with Ringer’s → mount in 1-2 drops of
1/3000 Neutral Red on the carry slide for 30min → rinse
with Ringer’s → dunk it with Ringer’s for 10min → cover it
with a piece of cover slide → observe the sample under
light microscope and draw them down.
 Observe the Golgi complex of the cat neural node and
draw them down.
 Results and discussion

1. Observations of the membrane vesicle system in

the chondrocytes from frog xiphoid bone
2. Observations of the Golgi apparatus in neurons
of the cat neural node
3. Structure properties and functions of the
membrane vesicle system, especially the Golgi
apparatus