inter Spem et Metum

When I suffer from protocol frustrations I would usually turn to Protocol Online for help. Besides the thousands of well-exploited protocols by labs from all over the world, there may be more than a thousand experienced & skilled bio-researchers--graduates, doctors & postdocs, who are willing to help--actually I was substantially helped recently.

It is not all about methodology or publications there. As common, people express their agony in work and hatred towards their bosses--of course no one was mentioned by real identity. It is alsp a common way that people try to seek amusement for their boring life. Luckily, it seems not so elusive...

Here I forward one thread by rotiprata in Lab Joke, originally post here.

Year 2003, a new chap joined our lab...usual practise...need blood...poke the freshies...
My friend went off to separate the blood to get the white blood cells while I prepared the microscope...
Moments later, he came in with a tube & made a slide with its content...
I looked under the microscope...
.....''erm....is this white blood cells?''.....
.....''of coz they are!''.....
.....''den y do they have long tails on them?''.....
.....''wat r u talking abt? let me see!''.....
.....''OH MY GOD! THESE ARE SPERMS!!!!''.....
.....''r u sure u took the correct sample???''.....
.....''OF COZ I DID! DAMN SURE!......
.....''U MEAN HIS BLOOD GOT SPERMS????!!!!''....
.....*dreaming of us goin on stage to present our remarkable finding on the international TV*.....i am getting very excited...
.....''OH MAN! I MUST TAKE PHOTOS OF THIS! I WAN MORE OF HIS BLOOD! HOW COME HIS BLOOD GOT SPERMS???!!!''....
......''dunno...but he sure need to find a gal & get married soon...its leaking into his blood circulation already...''
.........*click*....*click*....
.....''quick! make more slides! need to confirm! Boss must be damn proud of us for making such a discovery! I am goin to graduate soon! win lots of awards, maybe we can share a nobel prize huh??? we'll be famous!!!''...
.......*click**click**click*....
.....''erm....sherry???''.....
.....''YAR??? can u hurry with your slides? need to get more nicer pictures of sperm in blood, the sperm seems to be dying...''
.....''i got the wrong tube ah....''
''WAT!!!''
.......''yar...i forgot to label, so took the wrong one....this is mouse sperm....''
''WRONG TUUUBEEEEEEE??????!!!!!!''
Needless to say, he got a big scolding from me for not labeling his tubes.....after i regained my composure & started to delete the pictures...
.......''sherry?''......
......''WAT!?!?!?''......
.......''dun mention this to anybody ah?''......
......''OK LAH!''......

I kocked into this post the other day while I'm skimming over my Google Groups.I thought it was somewhat relative to my recent work so I replied in the thread.The next day I was informed of his reply indicated that I didn't hit. So I organized as much as I know to compose a second reply.And this seemde to satisfy him.

After re-read what I wrote,I was convinced that for sure there're still tons of flaws in my knowledge and comprehension although I didn't convince myself of what and where the flaws are.Anyhow I must know it if I want to do something in future.After all,illusion of knowledge is more dangerous than igorance of knowledge,as Barry Marshall,the 2005 Nobel laureate in Physiology or Medicine who visited our school recently,told in the lecture.

Well,I try to wrote that reply,in case that(dreaming that) I become a big wig and be asked to wrote a review or something.

Em...Realistically,consider it a review paper for a Master's diploma...


Here I post the thread.Sephy is me.


Mitotic spindle fiber stain
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From: Vincenzo Petix - view profile
Date: Sat, Apr 8 2006 5:27 am
Email: "Vincenzo Petix" <v.pe...@libero.it>
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Hi,
I'm a biologist who works in a genetics laboratory.
I'd like know if exists a method for stain mitotic spindle fibers
without use rabbits antibody
someone can help me? Thanks






From: Sephy - view profile
Date: Sun, Apr 9 2006 4:46 pm
Email: "Sephy" <phoenix...@gmail.com>
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Just search for the specific probe from Molecular Probes, devoured by
Invitrogen.

However, it would cost you about 500+ dollars...


To stain mitotic spindle fibers perhaps you'd better find a
tubulin-specifice probe.








From: Vincenzo Petix - view profile
Date: Sun, Apr 9 2006 6:23 pm
Email: "Vincenzo Petix" <v.pe...@libero.it>
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you didn't solve my problem ,but thank you very much for giving me your help

"Sephy" <phoenix...@gmail.com> ha scritto nel messaggio
news:1144572370.535129.102070@g10g2000cwb.googlegroups.com...



- Hide quoted text -
- Show quoted text -

> Just search for the specific probe from Molecular Probes, devoured by
> Invitrogen.

> However, it would cost you about 500+ dollars...


> To stain mitotic spindle fibers perhaps you'd better find a
> tubulin-specifice probe.









From: Sephy - view profile
Date: Mon, Apr 10 2006 4:37 pm
Email: "Sephy" <phoenix...@gmail.com>
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Uh...Well, maybe there are commonly 3 ways to stain microtubules as
well as other components of cytoskeleton like F-actins. It is the
purpose of you experiment that decides.

1.Stains with labelled probe,usually fluorescence labelled.It is
readily obtain from many vendors like Invitrogen or Sigma.In most
occasions,you can get a very nice photo from that.However it will cost
you much,and the standard protocols are not directly applicable-I
suffered from this recently so believe me.And the main drawback is you
can hardly stain a cell while it's living.So this method is eligable in
morpholgical research but unavailable when you want to do vital stain.


2.Immunofluorescence.Average cost,average effort required and quite
nice photos obtainable(by quite nice I mean it surely do some harm to
cells).There are tons of protocols about this method.I feel strange why
you aviod it.But it is you make decision.


3.GFP,recently GFP-mTn.As to label mitotic spindles,which mainly
contain microtubules,you could construct a fusion protein with
alpha-tubulin and GFP/GFP-mTn,express it in vivo,excite it,say cheese!
It's a perfect method because least harm,if not none,is done to
cells.Not only that,also there's no need trying to persuade your boss
to pay the bill for the kit,and you don't have to kill lovely bunnies
only to prepare antibodies.There are many articles on PubMed to which
you could refer.


Hope it would be useful.


I am now involved in a work on actin cytoskeletonin plant.Unfortuately
I am experiencing protocol frustrations.Fluorescence phalloidin from
Invitrogen sucks.The protocol enclosed is somewhat a piece of bullshit.

It's quite changable when warm breeze started to remove all of your thick garments.But I could sense an ominous feel this afternoon.And thst seem to be true.I am now blocked in the lab,raining cats&dogs outside.I don't wanna challenge cycling back only to get a whole change,so I just take this short time writing,hoping the pour won't last that long.

During the seminar the day before yesterday,I witnessed our boss:devil: pushed another girl into tears.By another I mean that was for the second time since I started the rotation-like work in the lab.

Strictly he didn't use any words,although I can tell the little bit threat within.However,it might be a common skill only for bosses:devil: to push his guys down.But sometimes I guess,that as in a position of P.I.,he is obliged byhimeself to be like that.

It is undoubtedly perfect that working under a boss both rich:wizard: and nice:cheers: ,but that will absolutely be a real rat race to be under that guy.If I am asked to make choice,I'd rather choose the former.Anyway,some of we students are very skilled in the battle with pervert bosses:devil: somehow,but hardly any could deal with poverty.

The boss:devil: had just returned from a conference trip to Italy.From there when he was dealing with his allowance,I guess,he just took out 1 or 2 Euro to buy a packet of chocolates,which he put out to us before seminar began.Well,another appropriate instance for that "Stick and Carrot" approach.

After all reports were given,he showed off photos of him taken in Milano and somewhere else.I was just impressed by three findings however:first was the incredible beauty of Milano,second was the clumsy skill of photography and last but not least,the nasty guy appeared in the center.

I am now beginnig to believe that dust to dust,all leaves to the root,and all rains drops love earth.Get going for a dance in the rain...