To calculate concentrations of pollen, diatoms, or other microfossils in samples, a spike of tracers is added to the sample during preparation. These tracers are typically plastic microspheres or
an exotic spore or pollen taxon. Of the latter, *Lycopodium* spores and *Eucalyptus* pollen have most commonly
been used. Microspheres are usually added as a suspension; while exotic spores or pollen are added either as a suspension or as tablets. A known quantity of tracers is added to the sample, and
during analysis the tracers are counted along with microfossils. The concentration of the microfossils is determined

In order to calculate concentration data, you must enter the necessary data in the spreadsheet. Tilia can calculate concentrations in three different ways:

Method 1:

- Quantity of sample (volume or mass)
- Quantity of spike medium added (volume of suspension or number of tablets)
- Concentration of spike medium (number of tracers per ml or tablet)
- Number of tracers counted during analysis.

Method 2:

- Quantity of sample (volume or mass)
- Quantity of spike (number of tracers) added to sample (usually calculated as the product of 2 and 3 in Method 1)
- Number of tracers counted during analysis

Method 3:

- Quantity of sample examined (A known quantity of the original sample is placed on a slide, and either the entire slide is counted or a known fraction of the slide is counted. Thus, concentration is directly calculated as (number of microfossils counted)/(quantity of sample examined.)

All variables added to the spreadsheet for calculating concentrations should be assigned to the CONC group. The codes and names of the CONC variables can be anything; however, if standard codes, as described below, are used, Tilia can automatically assign the CONC variables to the appropriate parameters described above.

The examples of CONC variables below are for different variations of Method 1, which is the most common method.

**Example with microsphere suspension:**

Code |
Name |
Element |
Units |
Group |

mic.susp:quantity added:ml |
Microsphere suspension |
quantity added |
ml |
CONC |

mic.susp:concentration:number/ml |
Microsphere suspension |
concentration |
number/ml |
CONC |

mic:counted:number |
Microspheres |
counted |
number |
CONC |

samp.quant:volume:ml |
Sample quantity |
volume |
ml |
CONC |

**Example with Lycopodium tablets; quantity is number of
tablets:**

Code |
Name |
Element |
Units |
Group |

Lyc.tab:quantity added:number |
Lycopodium tablets |
quantity added |
number |
CONC |

Lyc.tab:concentration:number/tablet |
Lycopodium tablets |
concentration |
number/tablet |
CONC |

Lyc.spik:counted:number |
Lycopodium spike |
counted |
number |
CONC |

samp.quant:volume:ml |
Sample quantity |
volume |
ml |
CONC |

**Example with Eucalyptus tablets; quantity is grams of tablets
added:**

Code |
Name |
Element |
Units |
Group |

Euc.tab:quantity added:g |
Eucalyptus tablets |
quantity added |
g |
CONC |

Euc.tab:concentration:number/g |
Eucalyptus tablets |
concentration |
number/g |
CONC |

Euc.spik:counted:number |
Eucalyptus spike |
counted |
number |
CONC |

samp.quant:volume:ml |
Sample quantity |
volume |
ml |
CONC |

**Example with Lycopodium tablets; quantity is number of tablets, sample quantity in
grams:**

Code |
Name |
Element |
Units |
Group |

Lyc.tab:quantity added:number |
Lycopodium tablets |
quantity added |
number |
CONC |

Lyc.tab:concentration:number/tablet |
Lycopodium tablets |
concentration |
number/tablet |
CONC |

Lyc.spik:counted:number |
Lycopodium spike |
counted |
number |
CONC |

samp.quant:mass:g |
Sample quantity |
mass |
g |
CONC |

Neotoma has default codes for taxa names and uses SI abbreviations for measurement units. However, Neotoma does not have default codes for elements and non-SI units, and, therefore, uses the complete element and measurement unit names in the codes. However, abbreviations can be used. Some sensible abbreviations are:

quantity added quant.add

concentration conc

number n

tablet tab

volume vol

**For example:**

Code |
Name |
Element |
Units |
Group |

Lyc.tab:quant.add:n |
Lycopodium tablets |
quantity added |
number |
CONC |

Lyc.tab:conc:n/tab |
Lycopodium tablets |
concentration |
number/ml |
CONC |

Lyc.spik:counted:n |
Lycopodium spike |
counted |
number |
CONC |

samp.quant:vol:ml |
Sample quantity |
volume |
ml |
CONC |

After the necessary CONC data are added to the spreadsheet, concentrations may be calculated in two different ways. The following examples will use Method 1 with a microsphere suspension entered in this spreadsheet:

Select either:

**Calc > Concentrations**

**Insert > Worksheet >
Concentrations**

Which will display this message dialog:

Selecting Yes will give the same result as Calc > Concentrations.

If No is selected, an empty Concentrations worksheet will appear, in which, for example, concentration results from Excel could be pasted.

**Calc > Concentrations** or
**Insert > Worksheet > Concentrations > Yes** will display the
Concentrations dialog box:

The **Methods** listed are those described above.

The **Groups to include** box
lists the taxa groups in the spreadsheet. Select the ones you wish to convert to concentrations. Click the
**Include All** button to include all groups.

On the right are boxes for the CONC variables in the spreadsheet. Select the appropriate variable in each box for the selected **Method**. Alternatively, if default CONC codes were used, click the **Default
Codes** button, and Tilia will attempt to select the appropriate variables.

The **Null Values**
box indicates whether null values (blank cells) in the spreadsheet should be treated as null (no data) or as zero
values.

**Include All** and **Default Codes** were checked, and null values are to be treated as zeros.

Click the **Ok** button to calculate concentrations, which will appear an a new worksheet.

The .tlx file used in this example may be downloaded from here (Gonzales & Grimm 2009).