Laboratory in Cellular Biology

GENERAL INFORMATION

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Attendance at laboratories is required. Absences must be justified. Laboratories start on time. Please arrive on time so that introductory

comments do not have to be repeated. 3 Students should wear laboratory coats to protect clothing. You are required to see that your own area is clean and tidy after each lab. 4 Before each laboratory you should review the general topic in your own notes and in any text-books you may have used- Biology (Fourth Edition), by Neil A. Campbell, .. You should also read the procedures in the Manual, and prepare, if necessary, a flow sheet of the sequence of steps in each experimental protocol before coming to the lab. There will rarely be time to read about a technique and practice it in the lab. 5 Eating and drinking are not permitted in the laboratory. Students are free to leave the laboratory for short periods when the experimental schedule allows. Students should always wash their hands before leaving the laboratory.

Remember, experiments are exciting to carry out, so your reports should be equally exciting to read!!!

Course Lectures and Outline:

In the first part of this course, we are going to study Basic chemical reactions, Carbon compound in living systems, Carbohydrates, Lipids, Proteins (here we learn about the extraordinary variety of protein structure and function that has evolved from permutations of just 20 amino acid building blocks). We continue the course with the Nucleic acid (molecules whose remarkable coding capacity puts them at the center of living systems). This chapter shows how just four nucleotides can be strung together in DNA to provide the information for specifying all of the hundreds of thousands of proteins in cells. We continue with Metabolism, Cytoplasmic organelles, Membrane structure and function. Here we learn about the complex cells with a nucleus and the simpler cells that lack one. In the second part we will study Respiration, Fermentation, Photosynthesis, and one of the central events of cell life, the division; Mitosis and Meiosis, Most likely, this part will be discussed in the lab. Session, rather than in the lecture. In the third part we will study Mendelians genetics, Chromosomes, Genetic material and Protein synthesis. Finally, we will study Genetics of viruses and bacteria, Control of gene expression and DNA technology. In the end, we should see that cells, proteins and genes are so interconnected that no part of biology can be effectively studied without reference to analyses at both the molecular and cellular levels of organization.

Introduction to Biology
Biology is the study of living beings at several levels. It deals with

fundamental questions such as the origin and evolution of plants and animals. The study of Biology also has vast practical applications. To many scientists have stressed the importance of the study of cell biology because the cell is the central unit of biological organization. There are millions of types of cells in the organisms on earth. All living creatures are made of cells. The simplest organisms are unicellular, while more complex organisms have multiple cells, as humans.

History of The cells Discovery

In 1858, Rudolf Verchow, a German pathologist, enunciated the cell theory: Every animal appears as the sum of vital units, each of which bears in itself the complete characteristics of life. It should be outlined that there are two components of the cell theory. It states: 1) that all living things are composed of cells and 2) that all cells arise from other cells. It means that all organisms are composed of cells, and all cells come from preexisting cells. Even viruses, which are not cells themselves, are entirely dependent on the presence and chemical machinery of cells for their reproduction. Living cells probably arose on earth about 3.5 billion years ago (3.5x109 years) by spontaneous reactions between molecules in an environment that was far from chemical equilibrium. All organisms, and all of the cells that

constitute them, are believed to descended from a common ancestor cell through evolution by natural selection. How can we define a cell?

A cell can be defined as a small membrane -bounded compartments filled with a concentrated aqueous solution of chemicals. The outer shell of cells, like

any shell, is built to hold the interior contents from leaking out into the surrounding environment. In order to maintain their integrity, cells need to be surrounded by an environment through which water cannot flow. A membrane compose of fatty molecules serves this purpose. The structure and function of those membranes will be discuss in chapter 5. There are many types of cells, could you give examples? There are plant cells, animal cells, eucaryotic cells, procaryotic cells and many others. Cells exhibit

considerable variation in properties based on different arrangements of components. Cells also vary in size, although most of them fall in the range of 5 to 20 m and their volumes from 1 to 1.000m3. Neurons have volumes that fit within this range, but they often have fine projections that may extend for meters. The simpler are prokaryotic cells or anuclear cells. Those single cells lack nuclear compartments and membrane bounded internal compartments. Prokaryotic cells evolved before others and are the bacteria of today. The other cells, eukaryotic cells, those that have a nucleus, are later products of evolution, and they display much more complicated modes of gene regulation, contain membrane-bounded nuclei and have other internal compartments (called organelles). Both kind of cells must do many things in order to survive. Cells must obtain and process energy, they must convert the genetic information of DNA into protein. What else characterize cells? The cell is autonomous: Under certain conditions, you can grow it in

isolation from other cells. We call this cell culture. Cells can live in the absence of the rest of the organism from which they were taken and thus are truly alive. Organisms only grow by the growth and division of cells.

Comparison of Procaryotic

&

Eucaryotic Organisms

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Prokaryotes

Eukaryotes

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Organisms Cell size

Bacteria and cyanobacteria generally 1 to 10m in linear dimension

protists, fungi, plants, and animals generally 5 to 100 m in linear dimension

Never forget, there is exceptions

Metabolism Organelles DNA anaerobic or aerobic few circular DNA in cytoplasm aerobic nucleus, mitochondria, chloroplasts, ER, etc. very long linear DNA containing many non coding regions: bounded by nuclear envelope RNA & prot. synthesized in same compartment RNA synthesized and processed in nucleus; proteins synthesized in cytoplasm Cytoplasm no cytoskeleton: cytoplasmic streaming endocytosis, and exocytosis absent Cell division chromosomes pulled apart by attachments to plasma membrane Cellular organization mainly unicellular cytoskeleton composed of protein filaments; cytoplasmic streaming; endo and exocytosis chromosomes pulled apart by cytoskeletal spindle apparatus mainly multicellular, with differentiation of many cell types

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Introduction to Biology

The cell is the central unit of biological organization. All living creatures are made of cells. The simplest organisms are unicellular. History of The cells Discovery

In 1858, Rudolf Verchow, enunciated the cell theory: 1) All living things are composed of cells 2) All cells arise from other cells. All organisms, and all of the cells that constitute them, are believed to descended from a common ancestor cell through evolution by natural selection. There are many types of cells: Cells also vary in size, The simpler are prokaryotic cells or anuclear cells. The other cells, eukaryotic cells, those that have a nucleus Organisms only grow by the growth and division of cells.

Laboratory Reports:
An essential feature of scientific investigation is a clear and concise report on the observations and measurements that have been made and the conclusions that have been derived from them. The conventional, and very logical, format of the published paper consists of a title, followed by a summary, then introduction, methods, results, discussion and references, usually in that order. For different reasons, I do not insist on full laboratory reports and suggest a shortened, or streamlined variation that may be followed in the weekly analysis of your experiments. The emphasis is placed on data handling and the conclusions drawn directly from the data; the theory underlying the experiment should be referenced to textbooks and reading assignments. The recommended form of the report should contain: Title, introduction, Methods, Results, discussion, Conclusions and references.

Title: A statement or sentence in general terms defining the experiment. It should be

on a separate page, which should also include the course title, your partners name, the date of the report.

Introduction:

An explanation of the objective and the significance of the

experiment, and the means whereby it will be achieved, I.e., a brief and general description of the design of the experiment. The introduction should total 100-150 words.

Methods: Detailed steps of the techniques are in the Manual and need not be
repeated, except for significant changes that are announced during the class.

Results: The original data, as collected, are included her together with the final
tables, graphs, etc. If the raw data is voluminous, it is best included as an appendix where it is available for arithmetical checks or further calculations. Ensure that data on

all variables that could influence the experiment are included e.g. temperature, elapsed time, concentrations, etc. Presentation of results is of paramount importance. Consider carefully how best to arrange the results so that they are immediately understandable to an informed reader. Graphs or tables should usually be usedthe former being of greatest value in displaying the relationships between fixed and dependent variables, while the latter may be more applicable if experimental conditions are being altered. Histograms can also be used for visual display of such data. Graphs and tables must be numbered and titled, and units are essential. In graphs, select scales that are convenient to read and to subdivide, and which fill the available space. If your experiment fails, or does not produce usable data, you may obtain data from another student in your section.

Discussion:

You must analyze as far as possible your data, giving possible

explanations for a failed experiment. Report both sets of data, and acknowledge the source of the data used for calculation.

Conclusions:

State briefly in words what you think is shown by the graphs or

tables. Show where it is consistent with expected results and with theory. Cite the appropriate theory by referring to specific pages, paragraphs or statements in textbooks, recommended readings and other articles. Point out deviations in the data, and discuss possible origins of these deviations. Consider sources of variability, and the statistical validity of the results. Finally, decide to what extent the results support or negate the original hypothesis stated in the introduction. You should aim to make your report concise and piercing, achieved by careful choice of words and phrases, and by rigorous editing. This section should be no longer than 1 page.

References: you should specify the complete source of any information you are
using in your lab. Report or you may have consulted (journals, articles, books, pages....).