Total iron: heme and non-heme ICP-MS

non-heme iron : chormagen colorimetric analysis Reference: Torrance JD, Bothwell TH. Tissue iron stores. Methods in hematology: iron, ed Cook JD (Churchill Livingstone, New York) 1980:90–115.

heme iron: Heme assay kit: DIHB-250 DIHM-250 biosystem

liable iron pool: calcein- acetoxymethyl ester staining

Of course, measurment of iron level related genes, such as mRNA levels of TFR1 or protein levels of Ferritin H/L is alternative method.

I dont know about the best way, but one way of monitoring intracellular iron availability is to measure the levels of certain iron-dependent transcripts, such as transferrin receptor (elevated under iron-starvation) and ferritin (repressed under iron-starvation).

For more info, check out:

https://www.researchgate.net/publication/51232830_An_optimal_method_of_iron_starvation_of_the_obligate_intracellular_pathogen_Chlamydia_trachomatis?ev=prf_pub

Article An Optimal Method of Iron Starvation of the Obligate Intrace...

Perl's iron stain is a common method for demonstrating iron in tissues.

One of the alternative methods to using iron chelator for quantification of intracellular iron content of cells will be using QuantiChromTM Iron Assay Kit (Bioassay Systems, DIFE-250) which involves preparing a cell lysate and measuring Optical density of the sample at 590 nm ( ICP-MS).

However if theres a preferance to using chelators, you can look at colorimetric methods to meaure intracellular iron content. For more information you can take a look at the following: Colorimetric ferrozine-based assay for the quantitation of iron in cultured cells.(Anal (Riemer J et al, Biochem. 2004 Aug 15;331(2):370-5.).

However, I highly recommend that you take a look at[27] Rapid Colorimetric Micromethod for the Quantitation of Complexed Iron in Biological Samples

By WAYNE W. FISH in METHODS IN ENZYMOLOGY, VOL. 158- 1988

here, you can find a very simple method in which uses a reagent to release complexed-iron and a chelator to bind the released iron and measure the difference in absorbance before and after using the chelator.

I do have a copy of this method, so if you are interested I will be more than glad to share it with you.

Good luck

Its already decades ago that I have been in that "business", however, I well remember that challenge. First, colourimetric assays always need a real iron free lab environment, second, it makes a difference where the iron is located, in ferritin, haemosiderin or elsewhere, third, it is a question what exactly you want to know. The total iron? Readily available iron? The enzymatic active iron? Intermediate chelates? Just from what I remember. Iron chelators? To me, even more difficult. Could you, please, specify this more directly? (Sorry to answer with a question, but this could trigger a discussion more in depth) Good success

Total iron: heme and non-heme ICP-MS

non-heme iron : chormagen colorimetric analysis Reference: Torrance JD, Bothwell TH. Tissue iron stores. Methods in hematology: iron, ed Cook JD (Churchill Livingstone, New York) 1980:90–115.

heme iron: Heme assay kit: DIHB-250 DIHM-250 biosystem

liable iron pool: calcein- acetoxymethyl ester staining

Of course, measurment of iron level related genes, such as mRNA levels of TFR1 or protein levels of Ferritin H/L is alternative method.

here is a method for labile iron, we use in our lab:

http://www.sciencedirect.com/science/article/pii/S0891584902010067

For the total Fe al AAS or AES methods are the best you can get.