need to measure the refractive index of immersion oil

[Kyle]

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Hi All:

 

I have a question about the refractive index of immersion oil. In my
experiment, I need to use the immersion oil as a control specimen to measure
the refractive index of other tissues. Here the immersion oil was slightly
stained for fluorescence. As the dye I used was very little amount, I
assumed that it would not alter the refractive index of the immersion oil.
However, this assumption must be verified as it is important to the accuracy
of the results.

 

With this background, I have two questions:

 

1. Does anybody have the similar experience? If yes, could you briefly
describe the situations you ever met?

 

2. If I want to experimentally measure the refractive index of the stained
immersion oil, what kind of instrument is proper to this work? I'm not
familiar with this kind of techniques and was completely confused after lots
of searching online.

 

Any suggestions or information will be appreciated.

 

All the best

 

Kuyu

 

[Alberto Diaspro]

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i would use one drop of the fluorescent oil close to one of the original and imaging with a good, Kohler adjusted, clean, phase contrast microscope
no way of distinguishing...same refractive index

Bianco, B.; Diaspro, A. & Tommasi, T. (1989), 'Quantitative image processing through phase laser-microscopy''Proceedings of SPIE', 1090, 44 47.
Beltrame, F.; Bianco, B.; Castellaro, G.; Diaspro, A. & Nicolini, C. (1985), 'Quantitative phase contrast optical microscopy of cells', Medical and biological Engineering and Computing 23, 263--264.

Il giorno 27/ago/2012, alle ore 07:06, Kuyu Wang <[hidden email]> ha scritto:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi All:
>
>
>
> I have a question about the refractive index of immersion oil. In my
> experiment, I need to use the immersion oil as a control specimen to measure
> the refractive index of other tissues. Here the immersion oil was slightly
> stained for fluorescence. As the dye I used was very little amount, I
> assumed that it would not alter the refractive index of the immersion oil.
> However, this assumption must be verified as it is important to the accuracy
> of the results.
>
>
>
> With this background, I have two questions:
>
>
>
> 1. Does anybody have the similar experience? If yes, could you briefly
> describe the situations you ever met?
>
>
>
> 2. If I want to experimentally measure the refractive index of the stained
> immersion oil, what kind of instrument is proper to this work? I'm not
> familiar with this kind of techniques and was completely confused after lots
> of searching online.
>
>
>
> Any suggestions or information will be appreciated.
>
>
>
> All the best
>
>
>
> Kuyu
>
>

[Littlejohn, George]

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Dear Kuyu,

To empirically measure refractive index I have used an Abbé Refractometer, waterbath and thermometer. It works well for fluids such as immersion oil.

Best wishes,
George

******************************
Dr. George Littlejohn
School of Biosciences,
University of Exeter,
Halpin Laboratory,
Geoffrey Pope Building,
Stocker Road,
EX4 4QD, UK
******************************
Tel:  [phone] (Lab.)
      [phone] (Office)
Fax:  [phone]
E-mail:  [hidden email]
******************************
http://www.illuminatedcell.com/improved-imaging.html
http://projects.exeter.ac.uk/ein/
http://clumpy.ex.ac.uk


On 27 Aug 2012, at 07:30, "Alberto Diaspro" <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> i would use one drop of the fluorescent oil close to one of the original and imaging with a good, Kohler adjusted, clean, phase contrast microscope
> no way of distinguishing...same refractive index
>
> Bianco, B.; Diaspro, A. & Tommasi, T. (1989), 'Quantitative image processing through phase laser-microscopy''Proceedings of SPIE', 1090, 44 47.
> Beltrame, F.; Bianco, B.; Castellaro, G.; Diaspro, A. & Nicolini, C. (1985), 'Quantitative phase contrast optical microscopy of cells', Medical and biological Engineering and Computing 23, 263--264.
>
> Il giorno 27/ago/2012, alle ore 07:06, Kuyu Wang <[hidden email]> ha scritto:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hi All:
>>
>>
>>
>> I have a question about the refractive index of immersion oil. In my
>> experiment, I need to use the immersion oil as a control specimen to measure
>> the refractive index of other tissues. Here the immersion oil was slightly
>> stained for fluorescence. As the dye I used was very little amount, I
>> assumed that it would not alter the refractive index of the immersion oil.
>> However, this assumption must be verified as it is important to the accuracy
>> of the results.
>>
>>
>>
>> With this background, I have two questions:
>>
>>
>>
>> 1. Does anybody have the similar experience? If yes, could you briefly
>> describe the situations you ever met?
>>
>>
>>
>> 2. If I want to experimentally measure the refractive index of the stained
>> immersion oil, what kind of instrument is proper to this work? I'm not
>> familiar with this kind of techniques and was completely confused after lots
>> of searching online.
>>
>>
>>
>> Any suggestions or information will be appreciated.
>>
>>
>>
>> All the best
>>
>>
>>
>> Kuyu
>>
>>

[Chen, De (NIH/NCI) [C]]

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Hi Kuyu:

You may be able to estimate the refractive index with the following formula:

n=n 1 V 1 +n 2 V 2 / V 1 +V 2

n is the refractive index of the mixtrure, n1, n2 are that of the respective solution, and V1, V2 are their respective volume. We used this method to get the right refractive index at 80 degrees.

De
________________________________________
From: Kuyu Wang [[hidden email]]
Sent: Monday, August 27, 2012 1:06 AM
To: [hidden email]
Subject: need to measure the refractive index of immersion oil

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Hi All:



I have a question about the refractive index of immersion oil. In my
experiment, I need to use the immersion oil as a control specimen to measure
the refractive index of other tissues. Here the immersion oil was slightly
stained for fluorescence. As the dye I used was very little amount, I
assumed that it would not alter the refractive index of the immersion oil.
However, this assumption must be verified as it is important to the accuracy
of the results.



With this background, I have two questions:



1. Does anybody have the similar experience? If yes, could you briefly
describe the situations you ever met?



2. If I want to experimentally measure the refractive index of the stained
immersion oil, what kind of instrument is proper to this work? I'm not
familiar with this kind of techniques and was completely confused after lots
of searching online.



Any suggestions or information will be appreciated.



All the best



Kuyu

[Johannes Helm]

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Dear Kuyu,

as already indicated by George Littlejohn, the Abbé refractometer is a
suitable tool for measuring refractive indices.

However, the Abbé refractometers, which I am familiar with, can be used on
one wavelength, only. Depending on the degree of precision and accuracy
you demand for your measurements, you might prefer to measure the
refractive indices at different wavelengths. For that purpose, you'd need
access to a Pulfrich or Michelson interferometer. Possibly, an optics
group at your local physics department could help you in gaining access to
that type of instrument.


Best wishes,
Johannes


> *****
> To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi All:
>
>
>
> I have a question about the refractive index of immersion oil. In my
experiment, I need to use the immersion oil as a control specimen to
measure
> the refractive index of other tissues. Here the immersion oil was
slightly
> stained for fluorescence. As the dye I used was very little amount, I
assumed that it would not alter the refractive index of the immersion
oil.
> However, this assumption must be verified as it is important to the
accuracy
> of the results.
>
>
>
> With this background, I have two questions:
>
>
>
> 1. Does anybody have the similar experience? If yes, could you briefly
describe the situations you ever met?
>
>
>
> 2. If I want to experimentally measure the refractive index of the
stained
> immersion oil, what kind of instrument is proper to this work? I'm not
familiar with this kind of techniques and was completely confused after
lots

> of searching online.
>
>
>
> Any suggestions or information will be appreciated.
>
>
>
> All the best
>
>
>
> Kuyu
>
>
>

--
P. Johannes Helm, M.Sc. PhD
Seniorengineer
CMBN
University of Oslo
Institute of Basic Medical Science
Department of Anatomy
Postboks 1105 - Blindern
NO-0317 Oslo

Voice: [phone]
Fax: [phone]

WWW: folk.uio.no/jhelm