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3 Graduate students learn the experimental work primarily through hands-on experience in the laboratory, and some find this learning process rather frustrating. So, what is the root of the problem, and how can young researchers make experiments to work effectively? 三人行，必有我師焉。 -- 孔夫子 This presentation summarizes what I have learned from many previous graduate students who have taught me to become a teacher.
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To hear is to forget;
To see is to remember;
To do is to know!
Everything you have heard from lectures you’ll forget soon.
Whatever you have learned by watching others you’ll remember for a while.
Carrying out a challenging task with your own efforts makes you a capable and confident person!
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Scientific Attitude and Methods
Chauying J. Jen ( 任卓穎 )
Inspired by Prof. T. T. Sun ( 孫同天 )
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Graduate students learn the experimental work primarily through hands-on experience in the laboratory, and some find this learning process rather frustrating. So, what is the root of the problem, and how can young researchers make experiments to work effectively?
三人行，必有我師焉。 -- 孔夫子This presentation summarizes what I have learned from many previous graduate students who have taught me to become a teacher.
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研究之道在眼高，在手低，在找尋真相。
眼高，才能看到重要的研究題材。手低，才能完成嚴謹的研究成果。
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What is Scientific Attitude?
Complete intellectual honesty ( 誠實面對問題 ) is essential in our scientific work.
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IT is nothing but applying the IMRAD principle all the time and everywhere.
What are Scientific Methods?
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達成目的不擇手段？ 達成目的「要」擇手段，要選擇最正確有效的方法！
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However, it must be reproducible and informative.
A successful experiment does not necessary generate data in support of the hypothesis.
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常失敗者的特質Unproductive experimenters could not find the right way to solve the problem. They could not get the techniques to work reliably and predictably either.
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實驗失敗了…“When you can not find the source of trouble, when you have asked advice and done everything (?) you can, repeat the experiment…repeat and repeat…”一試再試試不成，再試一次； 因為…『失敗是成功之母』 ?!
-- Is this a piece of good advice?
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Blind trust in authority ( 迷信權威 ) and intolerable margin of error ( 低估失誤的嚴重性 ) are the underlying causes of inadequate experimental design and poor outcome. -- T T Sun 孫同天
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What—textbook, Scientific American, review article, SCI article, thesis, meeting abstract, research handbook, instrument manual, reagent data sheet, catalog, scientific news, notebook ……
Who—Nobel laureate, famous scientist, thesis advisor, other professors, senior students, fellow students, close friends, salesmen, myself ……
What or who are authorities?%
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Examples of blind trust:1. Specific mouse strain, cell line,....2. Enzyme activity, antibody specificity,… (original vial, stock solution, storage, working solution, labeling …)3. Computer printout or digital display (Mean & SE, pH, CO2 level, ...)4. Pipetman setting, expression pattern,… 5. Protocols (from handbooks, papers, theses, senior students, other labs, …)6. Typographic errors (27.3 mg or 2.93 g, …)7. ………
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blind trust Failure Authorities necessary doubt Success
However, there is a balance point between blind trust and necessary doubt for each experiment.
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Experiments that can not be repeated in short time are the ones with very low level of tolerable margin of error. They will only work under most carefully designed and executed conditions.
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How to avoid making unnecessary mistakes ( 不必要的錯誤 ) in a laboratory?
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Believe it or not ( 信不信由你 )?!Much of whether your experiment will work or not is already determined before you touch the bench!
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Have you attended Prof. O. Smithies’s Nobel Lecture “Turning Pages” at NCKUMC on Oct. 18, 2008?
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Notebook is to be used before, during, and after carrying out an experiment. Each experimental record is a mini IMRAD paper. This record should be made each time an experiment is carried out.
Use a notebook ( 寫實驗紀錄簿 )!%
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Things to be recorded in a notebook should include everything related to this particular experiment: date, purpose, background (reference, information source, etc.), design (table, “N+(N-1)” rule), reagents/solutions (data sheet), protocol (flowchart, make remarks during the experiment), results (numbers/pictures, printouts, statistic analysis) and discussion.
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Write down the protocol in advance, preferably with your own hands, because during writing you can re-evaluate each step involved in the experiment. Besides, it is like carrying out a cost-free virtual experiment.
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Trouble shooting ( 解決問題之道 )When a complicated experiment fails, a) Check your notebook first;b) Identify possible causes with the help of checkpoints;c) Make a working hypothesis about possible mistakes for failure;d) Design simple experiments to verify your working hypothesis;e) Revise your protocols or methods accordingly.
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How to optimize the conditions—the “N + (N- 1)” rule
Time (hr) : 6, 12, 18, 24pH : 6.4, 6.9, 7.4, 7.9Ab (dilution) : 1/500, 1/1000, 1/2000FCS (%) : 0.1, 0.5, 1, 2
The number of total experiments needed= 4 +3 + 2 +3 = 12
Sun TT (2004) Nat Rev Mol Cell Biol5: 577-581
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If positive controls do not work or not included, the negative results are absolutely meaningless.
If negative controls do not work or not included, the positive results are absolutely meaningless.
Example
Always include controls to verify your experimental results.
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Examine the positive and negative controls before discussing the experimental group.
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Taken together, for effectively doing experiments, it is advisable 1. to plan ahead and check your setup and reagents first; 2. to determine at the beginning whether any effect is produced under extreme conditions, e.g., a massive dose;
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3. to follow the process of systematic elimination;4. to start with a modest preliminary experiment (pilot experiment, screening test);初步測試結果應具參考價值，但僅供參考！ One must switch to carefully designed formal experiments as soon as the pilot test indicates promising outcomes.
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5. to take the greatest care with the essential points of the technique (the principle, the critical points, the sensitivity and specificity limitations…); 6. to stick to the well-designed protocol and finish the experiment with your best efforts ( 精心規劃的 實驗決不半途更改 );
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7. to carefully record all experimental details in the notebook;
8. to assess experimental results by some objective measurements whenever possible;
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9. to keep in mind that each stage of investigation must be established beyond reasonable doubt before passing to the next stage;
10. to remember that any satisfactory experiment must be reproducible.
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Now you have done everything as suggested, then you shall…
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Stand where you are, with what you have;Do what you can, but never satisfied!
--quoted by 連加恩
You are not required to be the best; but you must do your best!
-- 陳定南 to his son Jack
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萬山不許一溪奔，攔得溪聲日夜喧；到得前頭山腳盡，堂堂溪水出前村。 -- 楊萬里
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Use a notebook to help yourself…
I Make thorough background check.M Design the experiments carefully
and test the system systematically — the N + (N’- 1) rule. R Pay attention to details when
carrying out a well-planned experiment to get reliable results.
AndD Consult your mentor or
experienced friends, and make your own
judgments.
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Research learning curvesIII
III
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